Abstract:
A microchip inspection system including: a microchip having at least a target substance and a reagent which is fluorescently labeled and is specifically combined with the target substance, wherein a reaction of the target substance and the reagent is performed and detecting a fluorescence intensity in a detected section of the microchip is performed; a microchip holder; a photo detection section; a reaction start device; and a control section for controlling a reaction start timing of the reaction start device and a detection timing of the fluorescence intensity before and after a reaction by the photo detection section, wherein the control section correlates a detection timing of the fluorescence intensity before the reaction by the photo detection section with the reaction start timing by the reaction start device.

Description:
RELATED APPLICATION 
       [0001]    This application is based on Japanese Patent Application No. 2007-016153 filed on Jan. 26, 2007 in Japan Patent Office, the entire content of which is hereby incorporated by reference 
       FIELD OF THE INVENTION 
       [0002]    The present invention relates to a microchip inspection system, a microchip inspection apparatus and a program. 
       DESCRIPTION OF THE RELATED ART 
       [0003]    In recent years, a micro total analysis system (μTAS), which mixes and reacts a plurality of solutions, detects and analyzes a state of a reaction on a microchip, into which a microfluidic pathways have been integrally formed, has attracted attention. 
         [0004]    In the μTAS, there are merits, such as little amount of a specimen, a short reaction time and little waste. When it is used for a medical field, a burden to a patient can be eased by lessening amount of specimens (blood, urine, wiping liquid), and lessening amount of a reagent can lower the cost of an inspection. Moreover, since there is little amount of a specimen and a reagent, a reaction time is greatly shortened and the increase in efficiency of an inspection can be attained. Furthermore, since the apparatus is small, it can also be installed into a small medical institution, and it quickly can be inspected without selecting a place to be set. 
         [0005]    In a microchip inspection system, a specimen and a reagent, which are stored in the microchip, are conveyed along a fluidic pathway by supplying a drive liquid to a microchip from a micropump. Thereby, the specimen and the reagent are mixed in the fluidic pathway and a reaction occurs. A reaction liquid is conveyed to a detected section in the microchip, and a detection of a concentration of a target substance in the reaction liquid is performed in a detection section. 
         [0006]    For example, international publication No. WO/2005/108571 discloses an example of a detection of a target gene, which uses a microchip, onto which a microfluidic pathway has been integrally formed. First, a substance which traps the target gene, is fixed in the detection section of the microchip in advance. Next, a specimen and a reagent which is used for amplification of the target gene are reacted to generate an amplified product. By this, in case when the target gene is contained in the specimen, the target gene amplified in the amplified product will exist. Next, the amplified target gene is denatured to a single strand. The substance for trapping the target gene fixed in the detected section is allowed to trap the target gene by supplying the specimen to the detected section. Next, a target gene and a DNA probe are combined by a hybridization reaction by supplying the DNA probe to be hybridized to the target gene of a single strand to the detected section. The DNA probe is fluorescently labeled in advance. Next, a gold colloid liquid which is combined with the DNA probe combined with the trapped target gene, is supplied to the detected section, and the gold colloid is combined with the DNA probe. 
         [0007]    Next, in order to remove the gold colloid, which has not been combined, from the detected section, a cleaning fluid is supplied to the detected section. And the target gene is detected by optically detecting the concentration of the gold colloid in the detected section. 
         [0008]    Moreover, Unexamined Japanese Patent Application Publication No. 2001-255328 discloses that in the detection of the target gene in a biochip, detecting a fluorescence intensity of a fluorescence emitted from a treatment liquid by irradiating an excitation light to the treatment liquid, in which the fluorescently labeled target gene and the DNA probe are hybridized. 
         [0009]    Moreover, Unexamined Japanese Patent Application Publication No. 2003-517591 discloses a cycling probe method as a technology being capable of detecting the target gene in high sensitivity. It is a procedure of generating many fluorescent labeling to a small number of target gene by forming the target gene into a molding die and cyclically repeating the hybridization to the target gene of the DNA probe and isolation. 
         [0010]    In the cycling probe method, a labeling of the DNA probe is made by a fluorochrome using fluorescence resonance energy transfer. The DNA probe, in a normal state, exists as a fluorescent substance, which is a donor, and a quencher, which is an acceptor, becoming a pair. Even though an excitation light is irradiated, the quencher absorbs the fluorescence emitted from the fluorescent substance and the donor&#39;s fluorescence is not generated. In a state where the DNA probe has caused the hybridization reaction with the target gene, binding between the fluorescent substance and the quencher is cut and the fluorescence of the fluorescent substance is emitted outside. In case when binding between the fluorescent substance and the quencher is cut, the DNA probe separates from the target gene. And the DNA probe in a normal state combines with the target gene, which became free, by the hybridization reaction again. Thus, by repeating the hybridization reaction, the DNA probe from which the quencher was cut is amplified and strong fluorescence intensity is obtained along with a progress of the reaction. 
         [0011]    When conducting the detection by the fluorescence intensity as described in the Unexamined Japanese Patent Application Publication No. 2001-255328, in order to perform a precise detection, it will be necessary to correct the fluorescence intensity after the reaction by measuring the fluorescence intensity before the hybridization reaction. 
         [0012]    Especially when using the cycling probe method, the fluorescence from the fluorescent substance should be absorbed by the quencher before the hybridization reaction. However, in fact, the fluorescence from the fluorescent substance, which was not absorbed by the quencher, appears as a weak fluorescence. For this reason, there is a large significance in measuring the fluorescence intensity before a reaction and correcting the fluorescence intensity after the reaction. 
       SUMMARY OF THE INVENTION 
       [0013]    The present invention is made based on such a requirement, and an object of the present invention is to provide a microchip inspection system, a microchip inspection apparatus and a computer readable memory storing program, which can execute a precise detection for a difference of the fluorescence intensity before and after the reaction. 
         [0014]    One aspect of the invention is to provide, a microchip inspection system comprising: a microchip for detecting fluorescence intensity in a detected section, the microchip having at least a target substance and a reagent which is specifically combined with the target substance and is fluorescently labeled wherein a reaction of the target substance and the reagent is performed; a microchip holder which can store the microchip; a photo detection section including a light receiving section for receiving fluorescence from the detected section and a light emitting section for irradiating the detected section with excitation light, the photo detection section being provided corresponding to the detected section of the microchip stored in the microchip holder; a reaction start device, which starts the reaction; and a control section for controlling a reaction start timing of the reaction start device and a detection timing of the fluorescence intensity before and after reaction by the photo detection section, wherein the control section matches a detection timing of the fluorescence intensity before reaction by the photo detection section and reaction start timing by the reaction start device. 
         [0015]    Another aspect of the invention is to provide, a microchip inspection apparatus comprising: a microchip holder which can store a microchip for detecting fluorescence intensity in a detected section, the microchip having at least a target substance and a reagent which is specifically combined with the target substance and is fluorescently labeled wherein a reaction of the target substance and the reagent is performed; a photo detection section including a light emitting section provided corresponding to a detected section of the microchip stored in a microchip holder and irradiating the detected section with excitation light and a light receiving section for receiving fluorescence from the detected section; a reaction start device, which starts the reaction; and a control section for controlling a reaction start timing of the reaction start device and a detection timing of the fluorescence intensity before and after reaction by the photo detection section, wherein the control section correlates a detection timing of the fluorescence intensity before reaction by the photo detection section with reaction start timing by the reaction start device. 
         [0016]    Another aspect of the invention is to provide, a computer readable medium storing program wherein the program comprises, storing at least a target substance and a reagent which is specifically combined with the target substance and is fluorescently labeled, at a detection section of a microchip stored in a microchip holder; starting the reaction of the target substance and the reagent by a reaction start device, which starts the reaction; detecting a fluorescence intensity before reaction by the photo detection section; detecting a fluorescence intensity after reaction by the photo detection section; and correlating a detection timing of the fluorescence intensity before reaction by the photo detection section and reaction start timing by the reaction start device. 
     
    
     
       BRIEF DESCRIPTION OF THE DRAWINGS 
         [0017]      FIG. 1  illustrates an outline view of an inspection apparatus using a microchip related to an embodiment of the present invention. 
           [0018]      FIG. 2  illustrates a block diagram of an inspection apparatus using a microchip related to an embodiment of the present invention. 
           [0019]      FIG. 3  ( a ) illustrates an upper surface drawing of a microchip  1 . 
           [0020]      FIG. 3  ( b ) illustrates a side view of a microchip of a microchip  1 . 
           [0021]      FIG. 3  ( c ) is an illustration showing a situation where a covering board  109  in  FIG. 3  ( a ) is removed. 
           [0022]      FIG. 4  is an illustration showing a main portion of a control configuration of an inspection apparatus using a microchip related to an embodiment of the present invention. 
           [0023]      FIG. 5  illustrates a flowchart of a detection control related to an embodiment of the present invention. 
       
    
    
     DESCRIPTION OF THE PREFERRED EMBODIMENT 
       [0024]    Hereafter, although an embodiment of the present invention is described based on drawings, it is an example and does not limit to an embodiment of the present invention. 
         [0025]    Here, a “microchip” and an “inspection system” perform a chemical operation and a biochemical reaction, such as mixing, separation, synthesis, and extraction of bio molecules, such as protein and nucleic acid, such as DNA and RNA, within a small chip, and further denote a system configured by combining with an apparatus, which detects a reaction result. 
       (Apparatus Configuration) 
       [0026]      FIG. 1  illustrates an outline view of an inspection apparatus using a microchip related to an embodiment of the present invention. An inspection apparatus  80  is an apparatus which automatically reacts a specimen and a reagent, which were injected into a microchip  1  in advance, and automatically outputs a reaction result. 
         [0027]    There are provided a loading slot  83  for inserting the microchip  1  into inside of the apparatus, a display section  84 , a memory card slot  85 , a print output slot  86 , an operation panel  87 , and an external I/O terminal  88  in a case  82  of an inspection apparatus  80 . 
         [0028]    An inspection person inserts the microchip  1  in a direction of an arrow in  FIG. 1 , and starts an inspection by operating the operation panel  87 . When starting an operation, as will be explained later, a fluorescent reaction is started in the microchip  1  in the inspection apparatus  80 , and an inspection result based on the detection result of the fluorescence is displayed on the display section  84 . By operation of the operation panel  87 , the inspection result can be outputted from the print output slot  86  as a print output, or can be stored into a memory card inserted into the memory card slot  85 . Moreover, data can be saved in a personal computer, for example, by using a LAN cable from the external I/O terminal  88 . The inspection person takes out the microchip  1  from the loading slot  83  after a completion of the inspection. 
         [0029]      FIG. 2  is a block diagram of the inspection apparatus  80  using the microchip related to an embodiment of the present invention. A situation where the microchip is inserted from the loading slot  83  shown in  FIG. 1 , and the completion of a setting is illustrated in  FIG. 2 . 
         [0030]    The inspection apparatus  80  includes a drive liquid tank  10  which stores a drive liquid  11  for sending the specimen and the reagent which were injected into the microchip  1  in advance, a micro pump  5  for supplying the drive liquid  11  to the microchip  1 , a leak packing  6  to connect the micro pump  5  and the microchip  1  so that the drive liquid  11  may not leak, a thermoregulation unit  3 , which adjust a temperature of a necessary portion of the microchip  1 , a chip pressing plate  2  for making a close contact between the thermoregulation unit  3  and the leak packing  6  so that the microchip  1  may not be shifted, a pressure plate drive section  21  for raising and lowering the chip pressing plate  2 , a regulation member  22  for precisely positioning the microchip  1  against the micro pump  5 , and a photo detection section  4 , which detects a reaction state of the specimen in the microchip  1  and the reagent. 
         [0031]    The chip pressure plate  2  has retracted upwards from the position shown in  FIG. 2  in the initial state. Thereby, an insertion and an extraction of the microchip  1  is possible in the direction of an arrow X, and an inspection person inserts the microchip  1  from the loading slot  83  (refer to  FIG. 1 ) until the microchip  1  contacts the regulation member  22 . Then, the chip pressure plate  2  is lowered by the pressure plate actuator  21 , and contacts the microchip  1 . The lower surface of the microchip  1  closely contacts the thermoregulation unit  3  and the leak packing  6 . Thereby, the setting of the microchip  1  is completed. The microchip holder of the present invention is configured by the regulation member  22 , the chip pressure plate  2 , the thermoregulation unit  3  and the leak packing  6 . Moreover, a heater  23  for promoting an amplification reaction of the target gene and a hybridization reaction, which will be described later, by heating the detected sections  125  and  126  (refer to  FIG. 3 ) of the set microchip  1 , is provided inside of the chip pressure plate  2 . The heater  23  is equivalent to the heating device of the present invention. 
         [0032]    The thermoregulation unit  3  has a Peltier device  31  on a surface facing to the microchip  1 . When the microchip  1  is set in the inspection apparatus  80 , the Peltier device  31  is arranged to closely contact the microchip  1 . The Peltier device  31  to keep the reagent from denaturing cools a portion in which the reagent is stored. 
         [0033]    A photo detection section  4  is configured by an excitation light sources  41  as a light emitting section of the present invention, such as LED, an excitation light filter  42  for limiting a wavelength zone of the excitation light emitted from the excitation light source  41 , a condenser lens  43  for forming the excitation light passed through the excitation light filter  42  into a beam spot, which matches the size, which covers the detection sections  125  and  126  (refer to  FIG. 3 ) of the microchip  1 , a dichroic mirror  44  for reflecting the excitation light passed through the condenser lens  43  to irradiate the excitation light to the detected sections  125  and  126  of the microchip  1 , and for passing the fluorescence from the detected sections  125  and  126  of the microchip  1 , which has been emitted by the excitation light, a light receiving lens  45  for guiding the fluorescence passed through the dichroic mirror  44  to a light receiving section  47 , a detection light filter  46  for limiting the wavelength zone of the fluorescence passed through the receiving lens  45  and a light receiving section  47  configured by a photodiode for receiving the fluorescence passed through the detection light filter  46 . 
         [0034]    The micro pump  5  comprises a pump room  52 , a piezo-electric element  51  for changing a capacity of the pump room  52 , a first reduced fluidic pathway  53  located in the microchip  1  side of the pump room  52 , and a second reduced fluidic pathway  54  located in the drive fluid tank  10  side of the pump room. The first reduced fluidic pathway  53  and the second reduced fluidic pathway  54  are the extorted narrow fluidic pathway, and the first reduced fluidic pathway  53  is a longer fluidic pathway than the second reduced fluidic pathway  54 . 
         [0035]    In case when conveying the drive liquid  11  to the forward direction (direction which goes to the microchip  1 ), it drives the piezo-electric element  51  first so that the volume of the pump room  52  may be rapidly decreased. Then, turbulence occurs in the second reduced fluidic pathway  54 , which is a short, reduced fluidic pathway, and the flow path resistance in the second reduced fluidic pathway  54  becomes relatively large compared to the first reduced fluidic pathway  53 , which is a long reduced fluidic pathway. Thereby, the drive liquid  11  in the pump room  52  is dominantly pushed out in the direction of the first reduced fluidic pathway  53 , and is conveyed. Next, the piezo-electric element  51  is driven so that the capacity of the pump room  52  is gradually increased. Then, drive liquid  11  will flow from the first diaphragm fluidic pathway  53  and the second reduced fluidic pathway  54  along with the increase in capacity in the pump room  52 . Since a length of the second reduced fluidic pathway  54  is shorter compared to the first reduced fluidic pathway  53  at this time, a flow path resistance of the second reduced fluidic pathway  54  becomes smaller than that of the first reduced fluidic pathway  53 , and the drive liquid  11  dominantly flows into the pump room  52  from the direction of the second reduced fluidic pathway  54 . When the piezo-electric element  51  repeats the above operation, the drive liquid  11  will be conveyed in a forward direction. 
         [0036]    On the other hand, first, in case when conveying the drive liquid  11  to an opposite direction (direction which heads to the drive fluid tank  10 ), the piezo-electric element  51  is driven so that the capacity of the pump room  52  is gradually decreased. Since the length of the second reduced fluidic pathway  54  is shorter compared to the first reduced fluidic pathway  53 , the flow path resistance of the second reduced fluidic pathway  54  becomes smaller compared to that of the first reduced fluidic pathway  53 . Thereby, the drive liquid  11  in the pump room  52  is dominantly pushed out in the direction of the second reduced fluidic pathway  54 , and is conveyed. Next, the piezo-electric element  51  is driven so that the capacity of the pump room  52  is rapidly increased. Then, the drive liquid  11  will flow in from the first reduced fluidic pathway  53  and the second reduced fluidic pathway  54  along with the increase in the capacity in the pump room  52 . At this time, turbulence occurs in the second reduced fluidic pathway  54 , which is a short reduced fluidic pathway, and the flow path resistance in the second reduced fluidic pathway  54  becomes relatively large compared to the first reduced fluidic pathway  53 , which is a long reduced fluidic pathway. Thereby, the drive liquid  11  dominantly flows into the pump room  52  from the direction of the first reduced fluidic pathway  53 . When the piezo-electric element  51  repeats the above operation, the drive liquid  11  will be liquid conveyed in the opposite direction. 
       (Configuration of Microchip) 
       [0037]      FIG. 3  illustrates a configuration of the microchip  1  of an embodiment of the present invention.  FIG. 3  ( a ) illustrates an upper surface view of the microchip  1 .  FIG. 3  ( b ) illustrates a side view of the microchip  1 .  FIG. 3  ( c ) is an illustration showing a situation where a covering board  109  in  FIG. 3  ( a ) is removed. An example of the configuration is shown and it is not limited to this. 
         [0038]    In  FIG. 3  ( a ), an arrow shows the insertion direction, into which the microchip  1  is inserted in the inspection apparatus  80 , and  FIG. 3  ( a ) illustrates the surface, which becomes the lower surface of the microchip  1  at the time of insertion.  FIG. 3  ( b ) illustrates a side view of the microchip  1 . 
         [0039]    As shown in  FIG. 3  ( b ), the microchip  1  comprises a groove formed board  108  and the covering board  109 , which covers groove formed board  108 . 
         [0040]    As shown in  FIG. 3  ( c ), the microfluidic pathway and the fluidic pathway element for mixing and reacting the specimen and the reagent on the microchip  1  are provided in the groove formed board  108 . In  FIG. 3  ( c ), an arrow schematically shows the microfluidic pathway and the quadrangle schematically shows the fluidic pathway element. 
         [0041]    The following fluidic pathway elements are provided on the microchip  1 . 
         [0042]    Drive liquid injection sections  110   a - 110   e  are injection sections for injecting the drive liquid  11  from the micropump. 
         [0043]    A specimen injection section  113  is an injection section for injecting the specimen into the microchip  1 . 
         [0044]    Downstream of the drive liquid injection sections  110   a - 110   e , there are provided respectively a specimen storage section  120  for storing the specimen, a positive control storage section  121  for storing the positive control reagent of the target gene, a negative control storage section  122  for storing the negative control reagent, an enzyme and a substrate storage section  123  for amplifying a target gene, and a primer and a fluorescently labeled DNA probe storage section  124 . Each reagent is stored in each storage section in advance. 
         [0045]    The positive control reagent and the negative control reagent are reagents for monitoring whether the inspection was conducted in a normal manner. 
         [0046]    The fluorescently labeled DNA probe, which is used by the cycling probe method, is modified with chimera oligonucleotide composed of RNA and DNA. One end is modified with the fluorescent substance and another end is modified with the quencher. In an intact state, although the fluorescent is not emitted with a fluorescence resonance energy transition phenomenon, in case when the fluorescently labeled DNA probe and the target gene cause a hybridization reaction, an RNA part will be cut and the fluorescent will be emitted. 
         [0047]    Each of above-mentioned storage sections oppose to the Peltier device  31 , when the microchip  1  is set in the inspection apparatus  80 , and cooled down so that the specimen or the reagent stored in each storage section is not denatured. 
         [0048]    In the downstream of the specimen storage section  120 , the positive control storage section  121 , the enzyme and the substrate storage section  123  and the primer and the fluorescently labeled DNA probe storage section  124 , there is provided a detected section  125  for performing the amplification reaction and the detection to the mixed-solution of the specimen and the positive control reagent. 
         [0049]    In the downstream of the specimen storage section  120 , the negative control storage section  122 , the enzyme and the substrate storage section  123 , the primer and the fluorescently labeled DNA probe storage section  124 , there is provided a detected section  126  for performing the amplification reaction and the detection to the mixed-solution of the specimen and the negative control reagent. 
         [0050]    When setting the microchip  1  in the inspection apparatus  80 , the detected sections  125  and  126  oppose to the heater  23  and heated for a promotion of the amplification. 
         [0051]    The detected sections  125  and  126 , and the window section  109   a  of the covering board  109  corresponding to the detected sections  125  and  126  comprise materials, such as transparent glass and resin, so that optical detection can be performed. 
         [0052]    The specimen and the flow of each reagent are explained. First, prior to conducting the inspection by the microchip  1 , the inspection person injects the specimen from the specimen injection section  113  using a syringe. The specimen injected from the specimen injection section  113  is stored in the specimen storage section  120  through the microfluidic pathway, which communicates with the specimen storage section  120 . 
         [0053]    Next, the microchip  1  into which the specimen was injected is inserted in the loading slot  83  of the inspection apparatus  80  shown in  FIG. 1  by the inspection person, and it is set as shown in  FIG. 2 . Based on this operation, it becomes possible to drive the micro pump  5  and to inject the drive liquid  11  from the drive liquid injection sections  110   a - 110   e.    
         [0054]    First, when the drive liquid  11  is injected from the drive liquid injection section  110   a , the specimen stored in the specimen storage section  120  will be pushed out through the microfluidic pathway which communicates therewith and the specimen will be sent into the detected sections  125  and  126 . 
         [0055]    Next, when the drive liquid  11  is injected from the drive liquid injection section  110   b , the positive control reagent (reagent with the same DNA arrangement portion as the target gene) stored in the positive control storage section  121  will be pushed out through the microfluidic pathway which communicates therewith. The positive control reagent is sent into the detected section  125 , and mixed with the previously liquid conveyed specimen. 
         [0056]    Next, when the drive liquid  11  is injected from the drive liquid injection section  110   c , the negative control reagent (for example, purified water) stored in the negative control storage section  122  will be pushed out through the microfluidic pathway which communicates therewith. The negative control reagent is sent into the detected section  126 , and mixed with the previously liquid conveyed specimen. 
         [0057]    Next, when the drive liquid  11  is injected from the drive liquid injection sections  110   d  and  110   e , the enzyme and the substrate, and the primer and the fluorescently labeled DNA probe are respectively sent into the detected sections  125  and  126  from each storage section of  123  and  124  along the microfluidic pathway, which is communicated therewith, and mixed with the mixed-solution of the previously liquid conveyed specimen and control liquid. 
         [0058]    Next, when the detected sections  125  and  126  are heated by the heater  23 , in each detected section, the amplification reaction of the target gene (and the positive control DNA), the hybridization reaction of an amplified product and a fluorescent probe, and the isolation reaction of a fluorescent substance and the quencher concurrently progress, and the reaction from amplification to the fluorescent substance generation is collectively performed at once. 
         [0059]    And it becomes possible to perform the photo detection by irradiating the excitation light from the excitation light source  41  of the photo detection section  4  at the detected sections  125  and  126 , and by receiving the fluorescent emitted from the detected sections  125  and  126  on the light receiving section  47 . 
         [0060]    In addition, it may be possible to perform the amplification reaction, the hybridization reaction, and the detection with a separate fluidic pathway element as a modification. Moreover, on the apparatus configuration, the position of the heater  23  or the Peltier device  31  may be somewhat changed, as long as those mechanisms are satisfied. 
         [0061]    Table 1 shows an example of a rule for conducting a comprehensive determination of the inspection based on the detection result of the detected sections  125  and  126  is conducted. 
         [0000]    
       
         
               
               
             
               
               
               
             
           
               
                 TABLE 1 
               
             
             
               
                   
               
               
                 Existence of fluorescence luminescence 
                   
               
             
          
           
               
                 Specimen and positive 
                 Specimen and negative 
                   
               
               
                 control reagent 
                 control reagent 
               
               
                 (Detected section 125) 
                 (Detected section 126) 
                 Judgment 
               
               
                   
               
               
                 Exist 
                 Exist 
                 Positive (with a 
               
               
                   
                   
                 target gene) 
               
               
                 Exist 
                 Non-exist 
                 Negative (without a 
               
               
                   
                   
                 target gene) 
               
               
                 Non-exist 
                 Exist 
                 Re-examination 
               
               
                 Non-exist 
                 Non-exist 
                 (Occurrence of mixing 
               
               
                   
                   
                 abnormality in a 
               
               
                   
                   
                 reagent or reaction 
               
               
                   
                   
                 inhibition) 
               
               
                   
               
             
          
         
       
     
         [0062]    The positive control causes the amplification reaction, which is equivalent to the target gene even only with the reagent alone, the hybridization reaction with the DNA probe, and the generation reaction of the fluorescent substance. The negative control does not cause the generation reaction of the fluorescent substance in the reagent alone. 
         [0063]    By performing the reaction and the detection to the mixed-solution, which is a mixture of those reagents and the specimen, the good-or-bad determination of the inspection result shown in Table 1 is attained. 
         [0064]    In case of a positive result, namely, when the target gene is contained in the specimen, the mixed-solution of the specimen and the positive control reagent, and the mixed-solution of the specimen and the negative control reagent generate the fluorescence luminescence. In case of a negative result, namely, when the target gene is not contained in the specimen, the mixed-solution of the specimen and the positive control reagent generates the fluorescence luminescence by the reaction of positive control, but the mixed-solution of the specimen and the negative control reagent does not generate the reaction, therefore, the fluorescence luminescence is not generated. These two cases can be treated as the inspection results to which the normal reactions have been performed. 
         [0065]    On the other hand, for example, in case when an inhibitory substance of the reaction has been mixed into the specimen, neither of the mixed-solution of the specimen and the positive control reagent nor of the mixed-solution of the specimen and the negative control reagent, generate the fluorescence luminescence. In case when there is the fluorescence luminescence in the mixed-solution of the specimen and the negative control reagent, and no fluorescence luminescence in mixed-solution of the specimen and the positive control reagent, abnormalities, such as inactivation of the reagent stored in the microchip  1 , can be taken for a consideration. It is possible to prompt re-examination as these two cases have the inspection results, which performed the unusual reaction 
       (Control Configuration) 
       [0066]      FIG. 4  is an illustration showing the main portion of the control configuration of the inspection apparatus using the microchip related to an embodiment of the invention. The main configuration factors related to control of the present invention are shown. 
         [0067]    Centering on a CPU  90 , which executes a control of the inspection apparatus  80  corresponding to a program, a ROM  92 , a RAM  93 , a nonvolatile memory  94 , the photo detection section  4 , the Peltier device  31 , the heater  23 , the display section  84  and the operation panel  87 , are reciprocally connected by a bus  91 . 
         [0068]    The ROM  92  stores various control programs and data, which are executed by the CPU  90 . 
         [0069]    The RAM  93  is utilized as a work area by the CPU  90 , and in case when the CPU  90  executes control, the RAM  93  temporarily stores necessary programs and data. 
         [0070]    The nonvolatile memory  94  stores the detection result from the photo detection section  4 . 
         [0071]    The CPU  90  executes control based on the program stored in ROM  92 . It functions as a control section of the present invention. 
         [0072]    Since the explanations about the photo detection section  4 , the Peltier device  31 , the heater  23 , the display section  84 , and the operation panel  87  are mentioned above, the explanations are omitted. 
       (Flow of Detection Control) 
       [0073]      FIG. 5  illustrates a flowchart of the detection control related to an embodiment of the present invention. An example of a case when the amplification reaction of the target gene by the cycling probe method, and the hybridization reaction of the target gene and the DNA probe are started by the heater  23  executing heating is explained. The heater  23  is equivalent to a reaction start device of the present invention. 
         [0074]    The CPU  90 , which executes a processing based on the detection control program stored in the ROM  92 , performs the detection control. Further, under an assumption that an inspection is started by the input of an inspection start from the operation panel  87  of the inspection apparatus  80 , and that each reagent with which chemical operation of the mixing has been conducted in the fluidic pathway of the microchip  1  has already been sent into detected sections  125  and  126 . 
         [0075]    First, the CPU  90  measures the fluorescence intensity before the reaction by the photo detection section  4  (STEP S 1 ). Therefore, it becomes possible to detect a weak fluorescence from the fluorescent substance, which was not absorbed by the quencher. 
         [0076]    Next, the CPU  90  determines whether a predetermined time T 1  (for example, several seconds) has passed (STEP S 2 ). 
         [0077]    When it is determined that the predetermined time T 1  has passed (STEP S 2 ; Yes), the CPU  90  heats the detected sections  125  and  126  with the heater  23  (STEP S 3 ). A temperature control is executed to control a temperature suitable for the amplification reaction of the target gene, and the hybridization reaction. 
         [0078]    When it is determined that the predetermined time T 1  has not passed (STEP S 2 ; No), the CPU  90  will stand by until the predetermined time T 1  passes. 
         [0079]    Next, the CPU  90  determines whether a predetermined time T 2  (for example, several minutes) has passed (STEP S 4 ). Accordingly, it is determined whether the hybridization reaction fully progressed by passage of the predetermined time T 2 . 
         [0080]    When it is determined that the predetermined time T 2  has passed (STEP S 4 ; Yes), the CPU  90  will measure the fluorescence intensity after the reaction by the photo detection section  4  (STEP S 5 ). 
         [0081]    When it is determined that the predetermined time T 2  has not passed (STEP S 4 ; No), the CPU  90  will stand by until the predetermined time T 2  passes. In the meantime, the heating is continued. 
         [0082]    Next, the CPU  90  corrects the fluorescence intensity after the reaction based on the fluorescence intensity before the reaction (STEP S 6 ). For example, the CPU  90  corrects by deducting the fluorescence intensity before the reaction from the fluorescence intensity after the reaction. The difference of the fluorescence intensity before and after the reaction can be measured accurately and efficiently for every microchip by the correction. 
         [0083]    Next, the CPU  90  displays the fluorescence intensity after the correction on the display section  84 , or saves the fluorescence intensity after the correction at the nonvolatile memory  94  (STEP S 7 ). Then, the flow ends. 
         [0084]    As mentioned above, according to this embodiment of the present invention, since the control is conducted by managing the time (the predetermined time T 1 ) from the detection timing before the reaction to the heating start, which is the reaction start timing, for example, the difference of the fluorescence intensity by the reaction can be measured accurately in case when the predetermined time T 1  is set up so that a timing of the detection before the reaction is conducted just before the reaction start. 
         [0085]    In this embodiment of the present invention, as for a measurement before the reaction, the fluorescence intensity is measured before heating by the heater  23 . However, depending on the reagent, the reaction may not progress for a while even when the heating has started. In such a case, the fluorescence intensity may be measured concurrent with heating, or before the reaction progresses after heating. Having the fluorescence intensity measured before the reaction progresses after heating, for example, an influence on the fluorescence intensity or the detection by the convection of the liquid by heating of the detected sections  125  and  126  can be removed. In this case, STEP S 1  and STEP S 3  can be interchanged, in  FIG. 5 . 
         [0086]    After the predetermined time T 1  passes from a start of a heating, it is necessary to control an execution of the detection before the reaction, in case when the fluorescence intensity greatly changes by temperature. In this case, in  FIG. 5 , what is necessary is to drive the micropump  5  mentioned above at STEP S 1 , and to measure the fluorescence intensity before the reaction (STEP S 3 ) after the predetermined time T 1 , when the liquid conveyance is completed, has passed (STEP S 2 ). 
         [0087]    In this embodiment of the present invention, a case where the reaction was started by heating was explained. However, the control of this invention can also be applied to a case where the reaction progresses only by mixing. In this case, the liquid conveyance of the target substance and/or the reagent starts the reaction. And, the mixed completion timing when a mixture of the target substance and the reagent by the liquid conveyance completes, and the detection timing of the fluorescence intensity before the reaction are correlated. Therefore, even if it is the reaction without temperature dependence, the difference of the fluorescence intensity before and after the reaction can be measured accurately and efficiently for every microchip. In this case, after the liquid conveyance of the DNA probe from the detection reagent storage section  124  to the detected sections  125  and  126  has completed, it is preferred to measure the fluorescence intensity before the reaction. Since the liquid amount becomes equivalent before and after the reaction when stored in the detected sections  125  and  126 , the fluorescence intensity can be measured more accurately. A determination of the completion of the liquid conveyance may be conducted by a prediction from the amount of liquid conveyance of the micropump  5 . A sensor, such as a liquid level sensor for detecting the liquid amount of the detected sections  125  and  126 , may be provided separately. In this case, the micropump  5  is equivalent to the reaction start device and the liquid conveyance device of the present invention. 
         [0088]    Like this embodiment of the present invention, applying control of the present invention to the system, in which the DNA arrangement detection reaction of the target gene by the cycling probe method having a tendency to generate the weak fluorescence is conducted, has a large significant on accurately conducting measurement of the difference of the fluorescence intensity before and after the reaction. The reaction by the cycling probe method progresses quickly. Therefore, it is important to determine the measurement timing in a consideration of the reaction, when accurately measuring the difference of the fluorescence intensity before and after the reaction. 
         [0089]    As mentioned above, according to the present invention, since the detection timing before the reaction is set to match with the reaction start timing, the difference of the fluorescence intensity before and after the reaction can be measured accurately, and a precise detection can be performed.