Patent Publication Number: US-2011076716-A1

Title: Bacteria analysis apparatus, bacteria analysis method and computer program product

Description:
FIELD OF THE INVENTION 
     The present invention relates to a bacteria analysis apparatus, a bacteria analysis method and a computer program. 
     BACKGROUND 
     In the past, as an apparatus for determining the kind of bacteria included in a sample, for example, an apparatus described in United States Patent Publication No. 2005/0079569 has been known. 
     In this apparatus, a particle measurement apparatus using a flow cytometer is used to determine the kind of bacteria included in a sample. In this apparatus, a first measurement specimen is prepared from a sample and a second measurement specimen is prepared from a sample and an alkaline solution. The first measurement specimen and the second measurement specimen are measured by the particle measurement apparatus. In addition, on the basis of the measurement result of the first measurement specimen and the measurement result of the second measurement specimen, whether bacteria are gram-positive bacteria or gram-negative bacteria is analyzed. In the above-mentioned United States Patent Publication No. 2005/0079569, it is described that an alkaline solution of about pH14 is used as the alkaline solution. 
     However, in the above-mentioned United States Patent Publication No. 2005/0079569, there is a problem in that it is necessary to introduce a highly corrosive strong alkaline solution of about pH14 into the apparatus. 
     SUMMARY 
     The scope of the present invention is defined solely by the appended claims, and is not affected to any degree by the statements within this summary. 
     A first aspect of the present invention is a bacteria analysis apparatus comprising: a specimen preparation section for preparing a first measurement specimen from a sample by using a first enzyme; a detecting section for detecting bacteria included in the first measurement specimen; and an information processing section for outputting information for supporting determination of kind of bacteria included in the sample on the basis of the detection result of the first measurement specimen. 
     A second aspect of the present invention is a bacteria analysis method comprising: preparing a first measurement specimen from a sample by using a first enzyme; detecting bacteria included in the first measurement specimen; and outputting information for supporting determination of kind of bacteria included in the sample on the basis of the detection result of the first measurement specimen. 
     A third aspect of the present invention is a computer program product comprising: a computer readable medium; and instructions, on the computer readable medium, adapted to enable a general purpose computer to perform operations, comprising: obtaining detection result of bacteria included in a first measurement specimen which is prepared from a sample by using a first enzyme; and outputting information for supporting determination of kind of bacteria included in the sample on the basis of the detection result of the bacteria of the first measurement specimen. 
    
    
     
       BRIEF DESCRIPTION OF THE DRAWINGS 
         FIG. 1  is a perspective view showing the overall configuration of a bacteria analysis apparatus according to a first embodiment of the invention. 
         FIG. 2  is a block diagram showing an analysis section of the bacteria analysis apparatus according to the first embodiment shown in  FIG. 1 . 
         FIG. 3  is a diagram schematically showing a specimen preparation section and an optical detection section of the bacteria analysis apparatus according to the first embodiment shown in  FIG. 1 . 
         FIG. 4  is a diagram showing the structure of the optical detection section of the bacteria analysis apparatus according to the first embodiment shown in  FIG. 1 . 
         FIG. 5  is a flowchart for explaining the analysis operation of the bacteria analysis apparatus according to the first embodiment shown in  FIG. 1 . 
         FIG. 6  is a flowchart for explaining a first interpretation process of the analysis operation of the bacteria analysis apparatus according to the first embodiment shown in  FIG. 5 . 
         FIG. 7  is a diagram showing a first interpretation result screen showing the interpretation result of the first interpretation process of the analysis operation of the bacteria analysis apparatus according to the first embodiment shown in  FIG. 1 . 
         FIG. 8  is a flowchart for explaining a second interpretation process of the analysis operation of the bacteria analysis apparatus according to the first embodiment shown in  FIG. 5 . 
         FIG. 9  is a diagram showing a comprehensive analysis result screen showing the interpretation result of a comprehensive analysis process of the analysis operation of the bacteria analysis apparatus according to the first embodiment shown in  FIG. 1 . 
         FIG. 10  is a flowchart for explaining the comprehensive analysis process of the analysis operation of the bacteria analysis apparatus according to the first embodiment shown in  FIG. 1 . 
         FIG. 11  is a diagram for explaining a bacteria kind determination principle using lysozyme. 
         FIG. 12  is a diagram showing a first interpretation result screen showing the interpretation result of a first interpretation process of the analysis operation of a bacteria analysis apparatus according to a modified example of the first embodiment shown in  FIG. 1 . 
         FIG. 13  is a diagram showing a comprehensive analysis result screen showing the interpretation result of a comprehensive analysis process of the analysis operation of the bacteria analysis apparatus according to the modified example of the first embodiment shown in  FIG. 1 . 
         FIG. 14  is a flowchart for explaining a comprehensive analysis process of the analysis operation of a bacteria analysis apparatus according to a second embodiment shown in  FIG. 1 . 
         FIG. 15  is a diagram for explaining a bacteria kind determination principle using lysostaphin. 
         FIG. 16  is a flowchart for explaining the analysis operation of a bacteria analysis apparatus according to a third embodiment of the invention. 
         FIG. 17  is a flowchart for explaining a second interpretation process of the analysis operation of the bacteria analysis apparatus according to the third embodiment of the invention. 
         FIG. 18  is a flowchart for explaining a comprehensive analysis process of the analysis operation of the bacteria analysis apparatus according to the third embodiment of the invention. 
         FIG. 19  is a graph showing the relationship between the concentration of lysozyme and the number of bacteria which are  Enterococcus faecalis.    
         FIG. 20A  is a graph showing the relationship between the concentration of lysostaphin and the number of bacteria which are Staphylococcus aureus. 
         FIG. 20B  is a graph showing the relationship between the concentration of lysostaphin and the number of bacteria which are  S. sciuri.    
     
    
    
     DETAILED DESCRIPTION OF THE EMBODIMENT 
     Hereinafter, embodiments embodying the invention will be described with reference to drawings. 
     First Embodiment 
     The overall configuration of a bacteria analysis apparatus  1  according to a first embodiment of the invention will be described with reference to  FIGS. 1 to 4 . 
     As shown in  FIG. 1 , the bacteria analysis apparatus  1  according to the first embodiment includes a measurement section  2  for measuring a sample by flow cytometry and an analysis section  3  for analyzing the measurement result of the measurement section  2 . The measurement section  2  includes a rack table  4  for transporting a sample rack  200  (test tube rack) which stores test tubes  100  containing a sample, a specimen preparation section  5  for preparing a measurement specimen from a sample or the like, an optical detection section  6  for detecting information of bacteria or formed elements in urine from a measurement specimen and a control section  7  for controlling the rack table  4 , specimen preparation section  5 , optical detection section  6  and the like. On the side surface of the chassis of the measurement section  2 , a support  22  is attached via an arm  21 . The analysis section  3  composed of a personal computer is installed on the support  22 . In addition, the measurement section  2  and the analysis section  3  are connected by a LAN. 
     The rack table  4  has a function of transporting a sample rack  200  up to a predetermined sample suction position. 
     At the sample suction position, a syringe pump (not shown) suctions the sample (urine) in a test tube  100  by using a suction tube  8  and the sample is dispensed to a measurement specimen preparation section  51  to be described later. 
     As shown in  FIG. 3 , the specimen preparation section  5  has a function of preparing a measurement specimen by mixing a sample with a predetermined liquid. In greater detail, the specimen preparation section  5  includes the measurement specimen preparation section  51 , a diluent holding section  52 , a dye solution holding section  53  and an enzyme holding section  54 . In the first embodiment, the enzyme holding section  54  holds lysozyme which is a cell wall lytic enzyme. The specimen preparation section  5  can prepare a specimen for bacteria detection by mixing a dye solution with a diluent in the measurement specimen preparation section  51  in which a sample has been dispensed via the suction tube  8 . In addition, the specimen preparation section  5  can prepare a first specimen for bacteria determination by mixing an enzyme (lysozyme), a dye solution and a diluent in the measurement specimen preparation section  51  in which a sample has been dispensed. The measurement specimens (the specimen for bacteria detection and first specimen for bacteria determination) prepared in the specimen preparation section  5  are introduced to a flow cell  61  of the optical detection section  6  to be described later. 
     In the optical detection section  6 , optical measurement is performed by flow cytometry. That is, in the optical detection section  6 , a fine flow in which a measurement specimen is enclosed in a sheath liquid (not shown) is formed in the flow cell  61  and the fine flow is irradiated with laser light. As a specified configuration, as shown in  FIG. 4 , the optical detection section  6  includes a light source  62  composed of a semiconductor laser, a condenser lens  63  for focusing the laser light irradiated from the light source  62  on the fine flow of the flow cell  61 , two condenser lenses  64  and  65 , a dichroic mirror  66 , a scattered light receiving section  67  composed of a photodiode, a scattered light receiving section  68  composed of a photomultiplier and a fluorescent light receiving section  69  composed of a photomultiplier. 
     The condenser lens  64  has a function of focusing forward-scattered light beams, from among light beams emitted from a measurement specimen (fine flow) in the flow cell  61  which are irradiated with laser light, on the scattered light receiving section  67 . The condenser lens  65  has a function of focusing side-scattered light beams and side fluorescent light beams, from among light beams emitted from a measurement specimen in the flow cell  61  which is irradiated with laser light, on the dichroic mirror  66 . The dichroic mirror  66  is configured to reflect side-scattered light beams toward the scattered light receiving section  68  and to transmit side fluorescent light beams through the fluorescent light receiving section  69 . 
     The scattered light receiving section  67 , scattered light receiving section  68  and fluorescent light receiving section  69  are configured to convert received light into electric signals. These electric signals reflect the shape, number and the like of bacteria included in a sample. That is, the optical detection section  6  can detect bacteria included in a measurement specimen (sample). These electric signals are transmitted to the analysis section  3  via the control section  7 . 
     The operation of preparing a measurement specimen by the specimen preparation section  5 , the operation of the measurement by the optical detection section  6  and the like are carried out automatically by operation of a magnetic valve, a driving section and the like (not shown) depending on the control of the control section  7  (microcomputer) of the measurement section  2 . 
     In addition, as shown in  FIGS. 1 and 2 , the analysis section  3  is composed of a computer mainly including a control section  31  (see  FIG. 2 ), a display section  32  and an input device  33 . 
     As shown in  FIG. 2 , the control section  31  mainly includes a CPU  311 , a ROM  312 , a RAM  313 , a hard disk  314 , a reading device  315 , an I/O interface  316 , a communication section  317  and an image output interface  318 . The CPU  311 , ROM  312 , RAM  313 , hard disk  314 , reading device  315 , I/O interface  316 , communication section  317  and image output interface  318  are connected by a bus  319 . 
     The CPU  311  can execute computer programs stored in the ROM  312  and computer programs loaded to the RAM  313 . When the CPU  311  executes a program  34   a  for analyzing the kind of bacteria, which will be described later, the computer functions as the analysis section  3 . 
     Here, in the first embodiment, the CPU  311  of the analysis section  3  is configured to interpret the measurement data which is received from the control section  7  of the measurement section  2  via the communication section  317  and determine the kind of bacteria which may be included in a sample (urine), and to output information for supporting the determination of the kind of bacteria included in the sample on the basis of the determination result. In greater detail, the CPU  311  is configured to create a scattergram relating to the bacteria included in the sample by interpreting data from the measurement section  2 . In addition, the CPU  311  is configured to calculate a numerical value corresponding to the number of bacteria included in a sample on the basis of the scattergram. The CPU  311  is configured to obtain the degree of influence of an enzyme with respect to bacteria included in a sample on the basis of the detection result (a scattergram, a numerical value corresponding to the number of bacteria and the like) of a specimen for bacteria detection which is prepared from the sample and the detection result of a first specimen for bacteria determination which is prepared from the sample and enzyme (lysozyme) and to output on the display section  32  information for supporting the determination of the kind of bacteria included in the sample on the basis of the degree of influence. 
     The ROM  312  is composed of a mask ROM, a PROM, an EPROM, an EEPROM or the like, and computer programs which are executed by the CPU  311  and data which are used in the execution of the programs are recorded therein. 
     The RAM  313  is composed of a SRAM, a DRAM or the like. The RAM  313  is used to read computer programs which are recorded in the ROM  312  and the hard disk  314 . In addition, the RAM is used as a work area of the CPU  311  when these computer programs are executed. 
     In the hard disk  314 , various computer programs for execution by the CPU  311 , such as an operating system and an application program, and data which are used to execute the computer programs, are installed. The program for analyzing the kind of bacteria according to the first embodiment is also installed in the hard disk  314 . 
     The reading device  315  is composed of a flexible disk drive, a CD-ROM drive, a DVD-ROM drive or the like and can read computer programs or data which are recorded in a portable recording medium  34 . In addition, the program  34   a  for analyzing the kind of bacteria is stored in the portable recording medium  34  and the computer can read the program  34   a  for analyzing the kind of bacteria from the portable recording medium  34  and install the program in the hard disk  314 . 
     The above-mentioned program  34   a  for analyzing the kind of bacteria is provided by the portable recording medium  34  and can be also provided from an external device, which is connected to the computer by an electric communication line (which may be wired or wireless) to communicate therewith, through the electric communication line. For example, the program  34   a  for analyzing the kind of bacteria is stored in the hard disk  314  of a server computer on the internet and the computer accesses the server computer to download the program  34   a  for analyzing the kind of bacteria and to install the program in the hard disk  314 . 
     Further, in the hard disk  314 , for example, an operating system is installed for providing a graphical user interface environment, such as Windows (registered trade name) which is made and distributed by Microsoft Corporation in America. In the following description, the program  34   a  for analyzing the kind of bacteria operates on the above-mentioned operating system. 
     The I/O interface  316  is composed of, for example, a serial interface such as USB, IEEE1394 or RS-232C, a parallel interface such as SCSI, IDE or IEEE1284, and an analog interface including a D/A converter and an A/D converter. The input device  33  is connected to the I/O interface  316  and a user uses the input device  33  so as to input data to the computer. Accordingly, the user can issue an instruction (input of a measurement order) to the analysis section  3  to perform the measurement by the measurement section  2 . 
     For example, the communication section  317  is an Ethernet (registered trade name) interface. The analysis section  3  is configured to receive measurement data from the measurement section  2  via the communication section  317 . In addition, the analysis section  3  can transmit a control signal to the measurement section  2  via the communication section  317 . 
     The image output interface  318  is connected to the display section  32  composed of an LCD or a CRT so as to output to the display section  32  a picture signal corresponding to image data provided from the CPU  311 . The display section  32  is configured to display an image (screen) in accordance with an input picture signal. 
     Next, the analysis operation of the bacteria analysis apparatus  1  according to the first embodiment will be described with reference to  FIGS. 5 to 10 . 
     First, a tester issues an analysis instruction to the analysis section  3 . Accordingly, the analysis is started in the measurement section  2 . In greater detail, in Step S 1 , the control section  7  controls the specimen preparation section  5  to prepare a specimen for bacteria detection. That is, by mixing a specimen (urine), a diluent and a dye solution, a specimen for bacteria detection is prepared. After that, the prepared specimen for bacteria detection is transported to the optical detection section  6 . 
     Next, in Step S 2 , the control section  7  measures the specimen for bacteria detection. In greater detail, the specimen for bacteria detection is made to flow to the flow cell  61  so as to form a fine flow and the flow (sample) is irradiated with laser light from the light source  62 . Forward-scattered light beams, side-scattered light beams and side fluorescent light beams from the sample irradiated with laser light are received by the scattered light receiving section  67 , scattered light receiving section  68  and fluorescent light receiving section  69 , respectively. In the respective scattered light receiving section  67 , scattered light receiving section  68  and fluorescent light receiving section  69 , the forward-scattered light beams, side-scattered light beams and side fluorescent light beams are converted into electric signals. These electric signals are converted into digital data in the control section  7  and subjected to predetermined waveform processing. The measurement result (data corresponding to the forward-scattered light beams, data corresponding to the side-scattered light beams and data corresponding to the side fluorescent light beams) of the specimen for bacteria detection prepared by such processes is transmitted to the analysis section  3 . 
     Next, in Step S 3 , the CPU  311  of the analysis section  3  performs a first interpretation process. That is, as shown in  FIG. 6 , the CPU  311  creates a scattergram S 1  of the specimen for bacteria detection on the basis of the data corresponding to the forward-scattered light beams and the data corresponding to the side fluorescent light beams in Step S 10 . The CPU  311  sets an area A where bacteria are present in the scattergram S 1  in Step S 11  and counts the number of dots included in the area A in Step S 12  to obtain an enumeration result B 1  of the specimen for bacteria detection. The number of dots (enumeration result B 1 ) is a value reflecting the number of bacteria included in the sample. In Step S 13 , the CPU  311  stores the area A and the enumeration result B 1  on the hard disk  314 . 
     In Step S 4 , the CPU  311  displays a first interpretation result on the display section  32  of the analysis section  3  (personal computer). In greater detail, as shown in  FIG. 7 , a first interpretation result screen C including the scattergram S 1  of the specimen for bacteria detection created in Step S 10  and the enumeration result B 1  of the specimen for bacteria detection is displayed on the display section  32 . In the scattergram S 1  of the first interpretation result screen C, a dot group D 1  showing a bacteria distribution situation and the area A surrounding the group D 1  are shown. In addition, when bacteria are detected as a result of the analysis, the effect thereof and a message E for prompting the execution of the measurement for determining the kind of the detected bacteria are displayed along with the first interpretation result screen C. 
     By referring to the first interpretation result screen C, the tester decides whether or not to perform the measurement for determining the kind of the detected bacteria. When performing second interpretation, the tester issues an instruction to the bacteria analysis apparatus  1  to perform the measurement by pressing a measurement instruction button (not shown) on the screen displayed on the display section  32  of the analysis section  3 . In addition, in Step S 5  of  FIG. 5 , the CPU  311  determines whether the measurement instruction button has been pressed or not. When the measurement instruction button has not been pressed, the analysis process of the bacteria analysis apparatus  1  ends. When the measurement instruction button has been pressed, the CPU  311  instructs the measurement section  2  to start the measurement. 
     When the instruction for the measurement is issued, the control section  7  of the measurement section  2  controls the specimen preparation section  5  in Step S 6  to prepare a first specimen for bacteria determination. That is, by mixing an enzyme (lysozyme) with the sample (urine), diluent and dye solution, a first specimen for bacteria determination is prepared. After that, the prepared first specimen for bacteria determination is transported to the optical detection section  6 . 
     Next, in Step S 7 , the control section  7  measures the first specimen for bacteria determination as in Step S 2 . The measurement result (data corresponding to forward-scattered light beams, data corresponding to side-scattered light beams and data corresponding to side fluorescent light beams) of the first specimen for bacteria determination is transmitted to the analysis section  3 . 
     Next, in Step S 8 , the CPU  311  interprets the measurement data of the first specimen for bacteria determination (second interpretation process). That is, as shown in  FIG. 8 , in Step S 14 , a scattergram S 2  of the first specimen for bacteria determination is created on the basis of the data corresponding to the forward-scattered light beams and the data corresponding to the side fluorescent light beams. In Step S 15 , the area A set in Step S 11  (see  FIG. 6 ) is read and set in the scattergram S 2 . In Step S 16 , the number of dots included in the area A in the scattergram S 2  is counted to obtain an enumeration result B 2  of the first specimen for bacteria determination. The number of dots (enumeration result B 2 ) is a value reflecting the number of bacteria included in the first specimen for bacteria determination. In Step S 17 , the enumeration result B 2  is stored on the hard disk  314 . 
     After that, in Step S 18 , a comprehensive analysis is performed. 
     In the comprehensive analysis, first, as shown in  FIG. 10 , the enumeration result B 1  of the specimen for bacteria detection and the enumeration result B 2  of the first specimen for bacteria determination stored in the hard disk  314  are read in Step S 19 . Next, in Step S 20 , a predetermined threshold which is set in advance is read from the hard disk  314 . 
     In Step S 21 , the CPU  311  determines whether or not the value of (enumeration result B 2 /enumeration result B 1 ) is equal to or less than the threshold. The (enumeration result B 2 /enumeration result B 1 ) is a value indicating the degree of influence of the lysozyme on the bacteria included in the sample. By comparing the degree of influence with the threshold, the degree of influence of the lysozyme is quantitatively determined. When the value of (enumeration result B 2 /enumeration result B 1 ) is equal to or less than the threshold, in Step S 22 , the CPU  311  determines that the bacteria included in the sample are gram-positive cocci which are not  Staphylococcus.  When the value of (enumeration result B 2 /enumeration result B 1 ) is greater than the threshold, in Step S 23 , the CPU  311  determines that the bacteria included in the sample are gram-negative bacilli or gram-positive cocci which are  Staphylococcus.    
     After the determination result is stored in Step S 24 , a comprehensive analysis result screen F showing the result of the comprehensive analysis is displayed on the display section  32  in Step S 9  (see  FIG. 5 ). As shown in  FIG. 9 , in the comprehensive analysis result screen F, the scattergram  51  displayed in the first interpretation result screen C in Step S 4 , the scattergram S 2  created in Step S 14  and information based on the determination result of the comprehensive analysis in the above-mentioned Step S 18  (Steps S 19  to S 24 ) are displayed. In the scattergram S 2 , a dot group D 2  and the area A which is the same as in the scattergram  51  are shown. In  FIG. 9 , an example is shown in which the enumeration result B 2  of the first specimen for bacteria determination is remarkably smaller than the enumeration result B 1  of the specimen for bacteria detection and the value of (enumeration result B 2  of the first specimen for bacteria detection/enumeration result B 1  of the specimen for bacteria detection) is equal to or less than a threshold. In this case, in Step S 22  (see  FIG. 10 ), the bacteria included in the sample are determined as gram-positive cocci which are not  Staphylococcus,  and in the comprehensive analysis result screen F, a message G suggesting that the detected bacteria are gram-positive cocci and a message H suggesting that the detected bacteria are not staphylococcal bacteria are displayed. By referring to the messages G and H, the kind of bacteria included in the sample can be determined. 
     Next, a bacteria kind determination principle using lysozyme will be described. 
     Regarding a sample including at least one of the following four kinds of bacteria, a specimen for bacteria detection is prepared from the sample, diluent and dye solution and a first specimen for bacteria determination is prepared from the sample, diluent, dye solution and lysozyme as in the analysis of the bacteria analysis apparatus  1  according to the first embodiment so as to be measured by flow cytometry, and scattergrams created by the measurement are shown in  FIG. 11 . The four kinds of bacteria are  Staphylococcus epidermidis  ( S. epidermidis ),  Staphylococcus aureus  ( S. aureus ),  Enterococcus faecalis  ( E. faecalis ) and  Escherichia coli  ( E. coli ). These four kinds of bacteria are known as bacteria which cause urinary tract infection. Particularly, an uncomplicated urinary tract infection is usually caused by anyone of the four kinds of bacteria.  Staphylococcus epidermidis  and  Staphylococcus aureus  are gram-positive bacteria which are  Staphylococcus. Enterococcus faecalis  is gram-positive bacteria of  Enterococcus  (which are not  Staphylococcus ).  Escherichia coli  are gram-negative bacilli. 
     As shown in  FIG. 11 , regarding  Staphylococcus  and  Escherichia coli,  a difference in the number of dots between the measurement result of the specimen for bacteria detection and the measurement result of the first specimen for bacteria determination cannot be shown. On the other hand, regarding Enterococcus faecalis, the number of dots of the measurement result of the first specimen for bacteria determination is remarkably smaller than in the measurement result of the specimen for bacteria detection. That is, it is found that lysozyme reacts only with  Enterococcus faecalis,  not with  Staphylococcus  and  Escherichia coli,  and carries out the bacteriolysis. 
     In this manner, a sample is divided into two to prepare a specimen for bacteria detection by using one sample without the addition of lysozyme and prepare a first specimen for bacteria determination by using the other sample with the addition of lysozyme. Whether or not the bacteria included in the sample are  Enterococcus faecalis  can be determined on the basis of the measurement results of the two measurement specimens. 
     As the concentration of lysozyme included in the first specimen for bacteria determination, it is desirable that in gram-positive bacteria which are not  Staphylococcus  such as  Enterococcus faecalis,  a concentration is appropriately set at which a difference in the number of dots between the measurement result of the specimen for bacteria detection and the measurement result of the first specimen for bacteria determination can be recognized. The concentration of lysozyme in the first specimen for bacteria determination is preferably in the range of 2.5 to 20 mg/mL. When the concentration of lysozyme is equal to or higher than 2.5 mg/mL, the difference in the number of dots between the measurement result of the specimen for bacteria detection and the measurement result of the first specimen for bacteria determination can be easily recognized. When the concentration of lysozyme is equal to or lower than 20 mg/mL, lysozyme can be easily dissolved in the first specimen for bacteria determination and thus the first specimen for bacteria determination can be easily prepared. 
     In this manner, a sample is divided into two aliquots to prepare a specimen for bacteria detection in one aliquot without addition of lysozyme and prepare a first specimen for bacteria determination in the other aliquot with addition of lysozyme and it can be determined whether or not the bacteria included in the sample are  Enterococcus faecalis  on the basis of the measurement results of the two measurement specimens. 
     In addition, in the configuration using lysozyme according to the first embodiment, it is difficult to perform a determination of bacteria which are not  Enterococcus faecalis  (determination of the bacteria analysis apparatus  1  between  Staphylococcus  and gram-negative bacilli). However, by using a difference between the pattern of the scattergram of  Staphylococcus  and the pattern of the scattergram of gram-negative bacilli,  Staphylococcus  and gram-negative bacilli can be determined. That is,  Staphylococcus  and gram-negative bacilli can be determined from the fact that in the pattern of  Staphylococcus,  the distribution state of dots has a steep slope (the pattern rises), and in the pattern of gram-negative bacilli, the distribution state of dots has a gradual slope (the pattern is flat). In this manner, on the basis of the analysis result of the bacteria analysis apparatus  1  according to the first embodiment and the shape of the patterns of the scattergrams,  Staphylococcus, Enterococcus faecalis  and gram-negative bacilli can be determined. 
     In the first embodiment, as mentioned above, by providing the analysis section  3  which outputs information for supporting the determination of the kind of bacteria included in a sample on the basis of the detection result of a specimen for bacteria detection prepared from the sample and the detection result of a first specimen for bacteria determination prepared from the sample and lysozyme, information for supporting the determination of the kind of bacteria included in a sample can be output by using an enzyme (lysozyme) with a low level of corrosion without introduction of a highly corrosive strong alkaline solution to the bacteria analysis apparatus  1 . 
     In the first embodiment, as mentioned above, the analysis section  3  obtains the degree of influence of lysozyme on the bacteria included in a sample on the basis of the detection result of a specimen for bacteria detection and the detection result of a first specimen for bacteria determination, and outputs information for supporting the determination of the kind of bacteria included in the sample on the basis of the degree of influence. Due to such configuration, by using lysozyme which is known to react with  Enterococcus faecalis  (gram-positive bacteria which are not  Staphylococcus ), it can be estimated that a sample in which the influence of lysozyme is recognized on the basis of the detection result of a specimen for bacteria detection and the detection result of a first specimen for bacteria determination includes the bacteria ( Enterococcus faecalis ). The kind of the estimated bacteria can be output as information for supporting the determination of the kind of bacteria included in the sample. 
     In the first embodiment, as mentioned above, the degree of influence of lysozyme on the bacteria included in a sample is obtained on the basis of the value reflecting the number of bacteria of a specimen for bacteria detection and the value reflecting the number of bacteria of a first specimen for bacteria determination, and outputs information for supporting the determination of the kind of bacteria included in the sample on the basis of the degree of influence. Due to such configuration, the degree of influence of lysozyme on the bacteria included in a sample can be easily obtained on the basis of the value reflecting the number of bacteria of a specimen for bacteria detection and the value reflecting the number of bacteria of a first specimen for bacteria determination. 
     In the first embodiment, as mentioned above, the analysis section  3  creates the scattergram S 1  relating to the bacteria included in a specimen for bacteria detection and the scattergram S 2  relating to the bacteria included in a first specimen for bacteria determination on the basis of the detection result of the specimen for bacteria detection and the detection result of the first specimen for bacteria determination, and obtains values (enumeration results B 1  and B 2 ) reflecting the number of bacteria included in the specimen for bacteria detection and the first specimen for bacteria determination on the basis of the scattergram S 1  and the scattergram S 2 . Due to such configuration, values reflecting the number of bacteria included in a specimen for bacteria detection and a first specimen for bacteria determination can be easily obtained on the basis of the scattergram S 1  and the scattergram S 2 . 
     In the first embodiment, as mentioned above, as information for supporting the determination of the kind of bacteria included in a sample, the name of bacteria which may be included in the sample is displayed on the display section  32 , and thus a tester can know the name of bacteria which may be included in the sample. Accordingly, it is possible to easily determine which bacteria are included in the sample. 
     In the first embodiment, as mentioned above, lysozyme reacts with gram-positive bacteria which are not  Staphylococcus.  Accordingly, by using lysozyme, a tester can easily determine whether the bacteria included in a sample are gram-positive bacteria which are not  Staphylococcus  or gram-negative bacteria and gram-positive bacteria which are  Staphylococcus.    
     In the first embodiment, as mentioned above, by analyzing urine by using the bacteria analysis apparatus  1 , the kind of bacteria included in the urine can be easily determined. 
     In the first embodiment, it is possible to determine whether or not staphylococcal bacteria are included in a sample. Staphylococcal bacteria may have drug resistance with respect to certain drugs. For example, when administering methicillin, staphylococcal bacteria mutate into methicillin-resistant  Staphylococcus aureus  in some cases. Accordingly, a determination of whether or not staphylococcal bacteria are included in a sample is important in the clinical tests. In the first embodiment, by referring to the messages G and H of the comprehensive analysis result screen F of the bacteria analysis apparatus  1 , it can be determined whether or not staphylococcal bacteria are included in a sample on the basis of the pattern (dot group D 1 ) of the scattergram S 1  of a specimen for bacteria detection and the pattern (dot group D 2 ) of the scattergram S 2  of a first specimen for bacteria determination. 
     In the above-mentioned first embodiment, an example has been shown in which the number of dots included in the area A in each of the scattergram S 1  of a specimen for bacteria detection and the scattergram S 2  of a first specimen for bacteria determination are counted. However, the invention is not limited thereto. As in a first interpretation result screen I according to a modified example of the first embodiment shown in  FIG. 12 , the number of all of the dots in a scattergram may be counted without defining the area A. 
     In the above-mentioned first embodiment, an example has been shown in which the scattergram S 1  of a specimen for bacteria detection and the scattergram S 2  of a first specimen for bacteria determination are displayed side by side in the comprehensive analysis result screen F. However, the invention is not limited thereto. As in a comprehensive analysis result screen J according to the modified example of the first embodiment shown in  FIG. 13 , the pattern (dot group D 1 ) of the scattergram S 1  of a specimen for bacteria detection and the pattern (dot group D 2 ) of the scattergram S 2  of a first specimen for bacteria determination may overlap with each other and be displayed in a scattergram K. In this manner, a tester can easily recognize a change in the pattern of the scattergram S 2  of the first specimen for bacteria determination with respect to the scattergram S 1  of the specimen for bacteria measurement. 
     In the above-mentioned first embodiment, an example has been shown in which the kind of bacteria is determined by comparing a ratio of the dot count result with a predetermined threshold. However, the invention is not limited thereto. The determination maybe carried out by comparing a proportion of the area where the pattern (dot group D 1 ) of the scattergram of a specimen for bacteria measurement and the pattern (dot group D 2 ) of the scattergram of a first specimen for bacteria determination overlap with each other with a predetermined threshold which is set in advance with respect to the overlapping proportion. The overlapping proportion of the patterns of the scattergrams is a value indicating the degree of influence of lysozyme on the bacteria included in a sample. When the degree of influence of lysozyme is quantified using the amount of pattern overlapping in this manner, it is not necessary to count the number of dots of the scattergram in the determination of the kind of bacteria. 
     In the above-mentioned first embodiment, an example of the message in the comprehensive analysis result screen has been shown. However, other messages maybe shown. For example, when it is determined that gram-positive bacteria ( Enterococcus faecalis ) which are not  Staphylococcus  are included, a message displaying the name of bacteria, such as “Staphylococcal bacteria are not included in the sample.  Streptococcus  may be included in the sample.” may be shown, or a message displaying the number of bacteria, such as “The number of staphylococcal bacteria included in the sample: 0” may be shown. 
     In addition, when it is determined that gram-negative bacilli or gram-positive cocci which are  Staphylococcus  are included, a message displaying the kind of bacteria, such as “Gram-negative bacteria may be included in the sample”, a message displaying the name of bacteria, such as “Staphylococcal bacteria or  Escherichia coli  may be included in the sample”, or a message displaying the number of bacteria, such as “The number of staphylococcal bacteria included in the sample: measurement failure” may be shown. 
     Second Embodiment 
     Next, a bacteria analysis apparatus  1  according to a second embodiment of the invention will be described with reference to  FIG. 14 . In the second embodiment, unlike the above-mentioned first embodiment in which lysozyme is used, an example using lysostaphin will be described. Since the structure of the bacteria analysis apparatus  1  according to the second embodiment is the same as in the above-mentioned first embodiment, except for an enzyme to be used and an interpretation process of the analysis section  3 , the description of the structure of the bacteria analysis apparatus will be omitted. Since the analysis flow and the interpretation flow are also the same as in the first embodiment, except for a comprehensive analysis process, the description thereof will be omitted. 
     In a comprehensive analysis process according to the second embodiment, in Steps S 119  and S 120  shown in  FIG. 14 , the same process is performed as in Steps S 19  and S 20  of  FIG. 10 . In Step S 121 , the CPU  311  determines whether or not the value of (enumeration result B 2 /enumeration result B 1 ) is equal to or less than a threshold. When the value of (enumeration result B 2 /enumeration result B 1 ) is equal to or less than a threshold, the CPU  311  determines that the bacteria included in a sample are gram-positive cocci which are  Staphylococcus  in Step S 122 . When the value of (enumeration result B 2 /enumeration result B 1 ) is greater than the threshold, the CPU  311  determines that the bacteria included in a sample are bacteria which are not gram-positive cocci which are Staphylococcus in Step S 123 . In Step S 124 , after the storage of the determination result, the comprehensive analysis result is displayed on the comprehensive analysis result screen of the display section  32 . 
     In the above-mentioned second embodiment, in the comprehensive analysis result screen, a scattergram of the detection result of a specimen for bacteria detection, a scattergram of the detection result of a first specimen for bacteria determination and information based on the determination result of the comprehensive analysis in S 122  and S 123  are displayed as in the above-mentioned first embodiment. In greater detail, when it is determined that the bacteria included in a sample are gram-positive bacteria which are  Staphylococcus,  a message such as “Staphylococcal bacteria are included in the sample” or “The number of staphylococcal bacteria included in the sample: 000” is displayed. When it is determined that the bacteria included in a sample are bacteria which are not gram-positive cocci which are Staphylococcus, a message such as “Staphylococcal bacteria are not included in the sample” or “The number of staphylococcal bacteria included in the sample: 0” is displayed. 
     Next, a bacteria kind determination principle using lysostaphin will be described. 
     As shown in  FIG. 15 , as in the case of lysozyme shown in  FIG. 11 , scattergrams of a measurement specimen which is prepared from a sample, a diluent and a dye solution and a measurement specimen which is prepared from a sample, a diluent, a dye solution and lysostaphin are created by flow cytometry. 
     As shown in  FIG. 15 , regarding  Enterococcus faecalis  and  Escherichia coli,  a difference in the number of dots between the measurement result of the specimen for bacteria detection and the measurement result of the first specimen for bacteria determination cannot be shown. On the other hand, regarding  Staphylococcus  ( Staphylococcus epidermidis  and  Staphylococcus aureus ), the number of dots of the measurement result of the first specimen for bacteria determination is remarkably smaller than in the measurement result of the specimen for bacteria detection. That is, it is found that lysostaphin reacts only with  Staphylococcus,  not with  Enterococcus faecalis  and  Escherichia coli.    
     In this manner, a sample is divided into two to prepare a measurement specimen by using one sample without the addition of lysostaphin and prepare a measurement specimen by using the other sample with the addition of lysostaphin. Whether or not the bacteria included in the sample are  Staphylococcus  can be determined on the basis of the measurement results of the two measurement specimens. 
     As the concentration of lysostaphin included in the first specimen for bacteria determination, it is desirable that in gram-positive cocci which are  Staphylococcus  such as  Staphylococcus aureus  and  Staphylococcus epidermidis,  a concentration is appropriately set at which a difference in the number of dots between the measurement result of the specimen for bacteria detection and the measurement result of the first specimen for bacteria determination can be recognized. The concentration of lysostaphin in the first specimen for bacteria determination is preferably in the range of 0.5 to 100 μg/mL. When the concentration of lysostaphin is in the above-mentioned range, the difference in the number of dots between the measurement result of the specimen for bacteria detection and the measurement result of the first specimen for bacteria determination can be easily recognized. The concentration of lysostaphin in the first specimen for bacteria determination is more preferably in the range of 0.5 to 2.5 μg/mL. At this time, the difference in the number of dots between the measurement result of the specimen for bacteria detection and the measurement result of the first specimen for bacteria determination can be more easily recognized, and thus the kind of bacteria included in the sample can be easily determined. 
     In this manner, a sample is divided into two aliquots to prepare a measurement specimen in one aliquot without addition of lysostaphin and prepare a measurement specimen in the other aliquot with addition of lysostaphin and it can be determined whether or not the bacteria included in the sample are  Staphylococcus  on the basis of the measurement results of the two measurement specimens. 
     In the second embodiment, as mentioned above, by determining the presence of reaction of bacteria by using lysostaphin which is a cell wall lytic enzyme specific for bacteria which are  Staphylococcus,  it can be easily determined whether or not the bacteria included in a sample are  Staphylococcus.    
     In the second embodiment, as mentioned above, since lysostaphin reacts with gram-positive bacteria which are Staphylococcus, a tester can easily determine whether the bacteria included in a sample are gram-positive bacteria which are  Staphylococcus  or bacteria which are not  Staphylococcus  by using lysostaphin. 
     Other effects of the second embodiment are the same as in the above-mentioned first embodiment. 
     Third Embodiment 
     Next, a bacteria analysis apparatus  1  according to a third embodiment of the invention will be described with reference to  FIGS. 16 and 18 . In the third embodiment, unlike the above-mentioned first and second embodiments, an example in which bacteria are analyzed by using two enzymes, that is, lysozyme and lysostaphin will be described. Since the structure of the bacteria analysis apparatus  1  according to the third embodiment is the same as in the above-mentioned first embodiment, except for an enzyme to be used and an interpretation process of the analysis section  3 , the description of the structure of the bacteria analysis apparatus  1  will be omitted. 
     In an analysis operation according to the third embodiment, first, in Steps S 201  to S 204  of  FIG. 16 , the CPU  311  performs the same process as in Steps S 1  to S 4  of  FIG. 5 . A tester decides whether or not to perform second interpretation by referring to a first interpretation result screen as in the above-mentioned first embodiment. When the second interpretation is performed, the tester instructs the bacteria analysis apparatus  1  to perform the second interpretation by pressing a measurement instruction button (not shown) on the screen displayed on the display section  32  of the analysis section  3 . In addition, in Step S 205 , the CPU  311  determines whether or not the measurement instruction button has been pressed. When the measurement instruction button has not been pressed, the analysis process of the bacteria analysis apparatus  1  ends. When the measurement instruction button has been pressed, the CPU  311  instructs the measurement section  2  to start the second interpretation. 
     When the instruction for the second interpretation is issued, in Steps S 206  and S 207 , the control section  7  of the measurement section  2  controls the specimen preparation section  5  to prepare a first specimen for bacteria determination and a second specimen for bacteria determination. That is, the first specimen for bacteria determination is prepared by mixing a sample (urine), a diluent, a dye solution and an enzyme (lysozyme) and the second specimen for bacteria determination is prepared by mixing a sample (urine), a diluent, a dye solution and an enzyme (lysostaphin). 
     Next, in Step S 208 , the first specimen for bacteria determination and the second specimen for bacteria determination are subjected to the detection (measurement). The measurement result (data corresponding to forward-scattered light beams, data corresponding to side-scattered light beams and data corresponding to side fluorescent light beams) is transmitted to the analysis section  3 . 
     Next, in Step S 209 , the second interpretation process is performed by the analysis section  3 . That is, in Step S 211  of  FIG. 17 , a scattergram of the detection result of the first specimen for bacteria determination is created on the basis of the data corresponding to forward-scattered light beams of the first specimen for bacteria determination and the data corresponding to side fluorescent light beams. In addition, in Step S 212 , the area (see the area A of  FIG. 7 ) set in the scattergram of the first interpretation is read and set in the scattergram of the first specimen for bacteria determination. In Step S 213 , the number of dots included in the area is counted in the scattergram of the first specimen for bacteria determination to obtain an enumeration result B 2 . The number of dots (enumeration result B 2 ) is a value reflecting the number of bacteria included in the first specimen for bacteria determination. In Step S 214 , the enumeration result is stored in the hard disk  314 . 
     In Steps S 215  to S 218 , as in Steps S 211  to S 214 , a scattergram of the detection result of the second specimen for bacteria determination is created, the number of dots included in the area is counted and an enumeration result B 3  of the dots is stored. 
     After that, in Step S 219 , comprehensive analysis is performed (see  FIG. 18 ). 
     In the comprehensive analysis, first, in Step S 220  of  FIG. 18 , the enumeration result B 1  of the specimen for bacteria detection and the enumeration result B 2  of the first specimen for bacteria determination stored on the hard disk  314  are read. Then, a predetermined threshold which is set in advance is read from the hard disk  314  in Step S 221 . 
     In Step S 222 , the CPU  311  determines whether or not the value of (enumeration result B 2 /enumeration result B 1 ) is equal to or less than the threshold. When the value of (enumeration result B 2 /enumeration result B 1 ) is equal to or less than the threshold, the CPU  311  determines that the bacteria included in the sample are gram-positive cocci which are not Staphylococcus in Step S 223 . After that, the determination result is stored in Step S 229 . 
     When the value of (enumeration result B 2 /enumeration result B 1 ) is greater than the threshold, the CPU  311  reads the enumeration result B 1  of the specimen for bacteria detection and the enumeration result B 3  of the second specimen for bacteria determination stored in the hard disk  314  in Step S 224 . Next, a predetermined threshold which is set in advance is read from the hard disk  314  in Step S 225 . In addition, in Step S 226 , it is determined whether or not the value of (enumeration result B 3 /enumeration result B 1 ) is equal to or less than the threshold. When the value of (enumeration result B 3 /enumeration result B 1 ) is equal to or less than the threshold, the CPU  311  determines that the bacteria included in the sample are gram-positive cocci which are  Staphylococcus  in Step S 227 . When the value of (enumeration result B 3 /enumeration result B 1 ) is greater than the threshold, it is determined that the bacteria included in the sample are gram-negative bacilli in Step S 228 . Here, in the above-mentioned Step S 222 , it has been determined that the bacteria included in the sample are not  Enterococcus faecalis  (the bacteria included in the sample are  Staphylococcus  or gram-negative bacilli), and thus when the value of (enumeration result B 3 /enumeration result B 1 ) is greater than the threshold (there are no effects of lysostaphin) in Step S 228 , it is estimated that the bacteria included in the sample are not  Enterococcus faecalis  and also not  Staphylococcus.  Accordingly, when the value of (enumeration result B 3 /enumeration result B 1 ) is greater than the threshold in Step S 228 , it can be determined that the bacteria included in the sample are gram-negative bacilli. 
     In Step S 229 , after the storage of the determination result, the comprehensive analysis result is displayed on the comprehensive analysis result screen of the display section  32  in Step S 210  (see  FIG. 16 ). In the comprehensive analysis result screen, the scattergram of the detection result of the specimen for bacteria detection, the scattergram of the detection result of the first specimen for bacteria determination, the scattergram of the detection result of the second specimen for bacteria determination and information based on the determination result in the above-mentioned Steps S 223 , S 227  and S 228  are displayed. The enumeration results B 1 , B 2  and B 3  are also displayed in the comprehensive analysis result screen. In greater detail, when it is determined that the bacteria included in the sample are gram-positive cocci which are not  Staphylococcus,  a message such as “Are  Streptococcus  included in the sample?”, “Staphylococcal bacteria are not included in the sample.  Streptococcus  may be included in the sample” or “The number of staphylococcal bacteria included in the sample: 0” is displayed. When it is determined that the bacteria included in the sample are gram-positive cocci which are  Staphylococcus,  a message such as “Are staphylococcal bacteria included in the sample?”, “Staphylococcal bacteria are included in the sample” or “The number of staphylococcal bacteria included in the sample: 000” is displayed. When it is determined that the bacteria included in the sample are gram-negative bacilli, a message such as “Are  Escherichia coli  included in the sample?”, “Staphylococcal bacteria are not included in the sample.  Escherichia coli  may be included in the sample” or “The number of staphylococcal bacteria included in the sample: 0” is displayed. 
     In the third embodiment, as mentioned above, by outputting information for supporting the determination of the kind of bacteria included in a sample on the basis of the detection result of a specimen for bacteria detection, the detection result of a first specimen for bacteria determination and the detection result of a second specimen for bacteria determination, the kind (bacteria which are  Staphylococcus,  gram-positive bacteria which are not  Staphylococcus  or gram-negative bacteria) of bacteria included in the sample can be more accurately estimated by using lysozyme and lysostaphin. 
     It should be considered that the disclosed embodiments are examples in all aspects but do not restrict the invention. The scope of the invention is defined with the claims, not with the above description of the embodiments, and all changes within the meaning and scope equivalent to the scope of the claims are included in the present invention. 
     For example, in the above-mentioned first to third embodiments, an example has been shown in which the kind of bacteria is determined by using lysozyme or lysostaphin. However, the invention is not limited thereto, and when there is an enzyme (cell membrane-digesting enzyme, cell wall lytic enzyme) reacting with certain bacteria, the determination may be performed by using the enzyme. By using such an enzyme, bacteria which are not the kinds of bacteria described in the above-mentioned first to third embodiments also can be determined. 
     In the above-mentioned first to third embodiments, an example has been described in which the sample is urine. However, the invention is not limited thereto, and another sample such as blood may be used if it can be measured by flow cytometry. 
     In the above-mentioned first to third embodiments, an example has been described in which the kind of bacteria is analyzed in the analysis section  3 . However, the invention is not limited thereto, and the analysis may be performed by the control section  7  of the measurement section  2 . 
     In the above-mentioned first to third embodiments, an example has been described in which a specimen for bacteria detection is prepared and measured by the bacteria analysis apparatus  1 . However, the invention is not limited thereto. A specimen for bacteria detection may be prepared and measured by a different apparatus, the obtained measurement result may be transmitted to the analysis section  3  of the bacteria analysis apparatus  1  and comprehensive analysis may be performed by using the above measurement result and the measurement result of a specimen for bacteria determination. At this time, the measurement result can be provided from an external device, which is connected to the analysis section  3  by an electric communication line (which may be wired or wireless) to communicate therewith, through the electric communication line. 
     In the above-mentioned first to third embodiments, an example has been described in which one measurement specimen preparation section  51  is provided. However, the invention is not limited thereto and a configuration may be provided in which at least two measurement specimen preparation sections are provided. For example, a configuration also may be provided in which two measurement specimen preparation sections are provided and prepare a specimen for bacteria detection and a first specimen for bacteria determination, respectively. 
     In the above-mentioned first to third embodiments, an example has been described in which one suction tube  8  is provided. However, the invention is not limited thereto and a configuration may be provided in which at least two suction tubes are provided. For example, a configuration also may be provided in which two measurement specimen preparation sections and two suction tubes are provided such that a specimen for bacteria detection and a first specimen for bacteria determination are dispensed to each of the measurement specimen preparation sections via each of the suction tubes. 
     In the above-mentioned first to third embodiments, an example has been described in which the degree of influence of an enzyme on the bacteria included in a sample is expressed by “enumeration result B 2 /enumeration result B 1 ”. However, the invention is not limited thereto. The degree of influence also may be expressed by, for example, “enumeration result B 2 -enumeration result B 1 ” or “enumeration result B 2 /(enumeration result B 1 +enumeration result B 2 )”. 
     In the above-mentioned first to third embodiments, an example has been described in which the display section  32  outputs the comprehensive analysis result. However, the invention is not limited thereto. The comprehensive analysis result maybe printed on paper or may be transmitted to another apparatus. 
     In the above-mentioned first to third embodiments, an example has been described in which when a tester instructs the second interpretation using an enzyme after the first interpretation without using an enzyme, the second interpretation is performed. However, the invention is not limited thereto and a configuration also may be provided in which the first interpretation and the second interpretation are automatically performed without an instruction of the tester. 
     In the above-mentioned first to third embodiments, an example has been described in which a specimen for bacteria detection is prepared and measured, the necessity of the second interpretation is decided by the measurement result, and a first specimen for bacteria determination is prepared and measured when the second interpretation is required. However, the invention is not limited thereto and a configuration also may be provided in which both of a specimen for bacteria detection and a first specimen for bacteria determination are prepared and measured regardless of the necessity of the second interpretation. 
     EXAMPLES  
     First Example 
     Examination of Optimum Concentration of Lysozyme 
     In order to examine the optimum concentration of lysozyme in a measurement specimen, the number of bacteria in the measurement specimen in which the concentration of lysozyme had been changed was measured by using the fully automated urine formed element analyzer UF-1000i (manufactured by SYSMEX CORPORATION). 
     (1) Preparation of Measurement Specimen 
     First,  Enterococcus faecalis  were added to bacterial broth and were left to cultivate overnight. With regard to the bacterial broth, a heart infusion medium (prepared by NISSUI PHARMACEUTICAL CO., LTD.) which is a liquid medium for general bacteria was used following the instruction manual, and after warming and melting, was then subjected to high-pressure steam sterilization. 
     Next, the bacterial broth subjected to the overnight cultivation was added to the heart infusion medium so as to dilute it at a ratio of 1/500 and then was stirred. This broth was incubated at 35° C. for 4 hours and was prepared as a  Streptococcus faecalis  specimen. 
     (2) Measurement of the Number of Bacteria 
     Lysozyme (prepared by Wako Pure Chemical Industries, Ltd.) was added to the above-mentioned  Streptococcus faecalis  specimen such that a final concentration was 10 mg/mL, and then a reaction was carried out at 37° C. for 5 minutes by a heat block. 
     The fully automated urine formed element analyzer UF-1000i (trade name) (manufactured by SYSMEX CORPORATION) was used to measure the number of bacteria included in the reacted specimen. UF II PACK-BAC (trade name) (prepared by SYSMEX CORPORATION) was used as a diluent and UF II SEARCH-BAC (trade name) (manufactured by SYSMEX CORPORATION) was used as a stain solution to measure the number of bacteria included in the specimen by following the instruction manual of UF-1000i. 
     Also in specimens to which lysozyme was added such that a final concentration was 0 mg/mL, 1 mg/mL, 2.5 mg/mL or 5 mg/mL, the number of bacteria was measured by using the same method as mentioned above. 
     (Result) 
       FIG. 19  shows a graph showing the number of bacteria at each concentration when the number of bacteria included in a specimen having a lysozyme concentration of 0 mg/mL was set to 100. 
     From the graph of  FIG. 19 , it was found that when a final concentration of lysozyme was equal to or higher than 2.5 mg/mL, the number of bacteria included in the specimen was smaller than in a specimen without the addition of lysozyme. In addition, it was found that the higher the final concentration of lysozyme, the smaller the number of bacteria included in the specimen. That is, it was found that the reaction between lysozyme and bacteria is dependent on the lysozyme concentration. From this, it was suggested that the higher the concentration of lysozyme included in a measurement specimen, the more easily the kind of bacteria can be determined. 
     Second Example 
     Examination of Optimum Concentration of Lysostaphin 
     In order to examine the optimum concentration of lysostaphin in a measurement specimen, the number of bacteria in the measurement specimen in which the concentration of lysostaphin had been changed was measured by using the fully automated urine formed element analyzer UF-1000i (trade name) (manufactured by SYSMEX CORPORATION). 
     (1) Preparation of Measurement Specimen 
     First,  Staphylococcus aureus  were added to bacterial broth and were left to cultivate overnight. With regard to the bacterial broth, a heart infusion medium (prepared by NISSUI PHARMACEUTICAL CO., LTD.) which is a liquid medium for general bacteria was used following the instruction manual, and after warming and melting, was then subjected to high-pressure steam sterilization. 
     Next, the bacterial broth subjected to the overnight cultivation was added to the heart infusion medium so as to dilute it at a ratio of 1/1000 and then was stirred. This broth was cultivated at 35° C. for 4 hours and was prepared as a  Staphylococcus aureus  specimen. 
     In addition, in place of the above-mentioned Staphylococcus aureus,  S. Sciuri  was used to prepare a  S. Sciuri  specimen by using the same method as mentioned above. 
     (2) Measurement of the Number of Bacteria 
     Lysostaphin (prepared by Wako Pure Chemical Industries, Ltd.) was added to the above-mentioned  Staphylococcus aureus  specimen such that a final concentration was 1.0 μg/mL, and then a reaction was carried out at 37° C. for 5 minutes by a heat block. 
     The fully automated urine formed element analyzer UF-1000i (trade name) (manufactured by SYSMEX CORPORATION) was used to measure the number of bacteria included in the reacted specimen. UF II PACK-BAC (trade name) (prepared by SYSMEX CORPORATION) was used as a diluent and UF II SEARCH-BAC (trade name) (manufactured by SYSMEX CORPORATION) was used as a stain solution to measure the number of bacteria included in the specimen by following the instruction manual of UF-1000i. 
     Also in specimens to which lysostaphin was added such that a final concentration was 0 μg/mL, 0.5 μg/mL, 2.5 μg/mL, 5.0 μg/mL, 10.0 μg/mL or 100.0 μg/mL, the number of bacteria was measured by using the same method as mentioned above. 
     Also in the  S. Sciuri  specimen, the measurement was performed by using the same method as mentioned above. 
     (Result) 
       FIG. 20A  and  FIG. 20B  shows a graph showing the number of bacteria at each concentration when the number of bacteria included in a specimen having a lysostaphin concentration of 0 μg/mL was set to 100. 
     From the graph of  FIG. 20A  and  FIG. 20B , it was found that in any case of the  Staphylococcus aureus  specimen and the  S. Sciuri  specimen, the number of bacteria in the specimen having a final lysostaphin concentration of 0.5 to 100 μg/mL was smaller than in the case in which lysostaphin was not added. In addition, it was found that when a final concentration of lysostaphin was in the range of 0.5 to 2.5 μg/mL, the reactivity between lysostaphin and bacteria was the highest and the number of bacteria was nearly zero. That is, it was found that the reaction between lysostaphin and the  Staphylococcus aureus  specimen and  S. Sciuri  specimen is not dependent on the concentration in the same manner as the reaction of lysozyme and there is an optimum concentration for the reaction. 
     From this, it was suggested that when a final concentration of lysostaphin is in the range of 0.5 to 100 μg/mL, the kind of bacteria included in a measurement specimen can be easily determined, and further, when a final concentration of lysostaphin is in the range of 0.5 to 2.5 μg/mL, the kind of bacteria can be more easily determined.