Abstract:
A testing kit to determine the presence of microorganisms in aqueous solutions and suspensions by means of using a hand held plastic bag containing at least one reaction chamber, wherein the kit is typically embodied with of plurality of reaction chambers, and wherein each of the reaction chambers may be differently embodied yet still prevent cross contamination between the plurality of reaction chambers contained in the plastic bag.

Description:
BACKGROUND OF THE INVENTION  
         [0001]    The present invention relates to testing kits designed to be used to test for the presence of microorganisms in aqueous solutions and suspensions. Water, one of society&#39;s most important resources, is essential for the life of humans and animals. At times, however, water and foods containing deleterious microorganisms can be hazardous.  
           [0002]    Many people every year are sickened, or worse yet, die due to consuming water and foods contaminated with microorganisms such as  E. coli  and salmonella. This is true for people who rely on well water as their source of drinking water, as well as for as people who drink water provided by a public utility. People are even sickened by playing in recreational waters containing harmful microorganisms. Hazardous microorganisms are a constant threat to humans and animals alike.  
           [0003]    Present testing kits which test for microorganisms have numerous shortcomings. They test for one type of microorganism, while potentially missing many others. This gives a false sense of security to someone about to consume the suspect water or food material which has been tested with one of these devices and given the “all safe” signal. The individual might think the water or food is free of  E. coli  since the test detected no presence of this bacteria, while at the same time the water is loaded with salmonella bacteria. Further, some of the present devices are complicated and expensive to manufacture.  
           [0004]    There is a need for a reliable, inexpensive, easy to use testing kit for detecting the presence of either a single or a plurality of microorganisms.  
         SUMMARY OF THE INVENTION  
         [0005]    A testing kit to determine the presence of microorganisms in aqueous solutions, and to determine the presence of microorganisms in suspensions. The testing kit has a hand held apparatus containing at least one reaction chamber. Each reaction chamber contains a reactive agent to test for a predetermined microorganism or group of microorganisms. Each reaction chamber also prevents cross contamination between the plurality of reaction chambers, if any, contained in the plastic bag. 
       
    
    
     BRIEF DESCRIPTION OF THE FIGURES  
       [0006]    [0006]FIG. 1 shows a perspective view of an embodiment of the testing kit.  
         [0007]    [0007]FIG. 2 shows a top plan of the testing kit of FIG. FIG. 3 shows a sectional view of the testing kit taken along line A-A of FIG. 1.  
         [0008]    [0008]FIG. 4 shows a perspective view of the testing kit with the container loaded with the aqueous solution to be tested.  
         [0009]    [0009]FIG. 5 shows a perspective view of the rupturable membrane reaction chamber prior to the rupturable membrane being ruptured.  
         [0010]    [0010]FIG. 6 show a perspective view of pressure being applied to the rupturable membrane reaction chamber shown in FIG. 5.  
         [0011]    [0011]FIG. 7 shows a perspective view of the rupturable membrane reaction chamber of FIG. 6 drawing in the aqueous solution to be tested, or suspension to be tested, or both.  
         [0012]    [0012]FIG. 8 shows a perspective view of the resealable aperture reaction chamber.  
         [0013]    [0013]FIG. 9 shows a perspective view of pressure being applied to the resealable aperture reaction chamber of FIG. 8.  
         [0014]    [0014]FIG. 9A shows a perspective view of the resealable aperture reaction chamber of FIG. 9 drawing the aqueous solution to be tested, suspension to be tested, or both. 
     
    
     DETAILED DESCRIPTION  
       [0015]    Definitions  
         [0016]    1. Microorganisms  110  include bacteria, the coliform group of bacteria, the  E. coli  group of bacteria, salmonella bacteria, listeria, fungi, yeasts, molds (which are taxonomically fungi and yeasts), viruses, fecal streptococcus, enterococcus, iron bacteria, sulphur bacteria,  Vibro cholerae,  and other microorganisms  110  well known to those skilled in the art.  
         [0017]    2. Reactive Agent  24  includes chromogens, chromogenic substrates, and chromogenic reactive agents. Reactive agents  24  may change color when mixed with microorganisms  110 , or materials excreted by microorganisms. Reactive agents are well known to those skilled in the art.  
         [0018]    3. Test Mixture  56  includes the aqueous solution to be tested  20 , the suspension to be tested  100 , or a mixture of the solution  20  and suspension  100  with the reactive agent  24 .  
         [0019]    [0019]FIG. 1 shows a side perspective view of the testing kit  10 . The testing kit  10  is a container  12  which may be embodied in the form of a plastic bag  13  defining a bag interior  17  therein, as show in FIGS.  1 - 4 . The plastic bag  13  allows the entire testing kit to be compactly contained, folded, and stored. The plastic bag  13  also allows the user to visually observe what is occurring in the bag interior  17 . The plastic bag  13  has a resealable opening  14  along an edge  16  thereof. That is, it may be repeatedly opened and closed by pulling the resealable opening  14  apart, and then compressing and or folding the resealable opening  14  together again. This type of plastic bag  13  is well known to those skilled in the art. FIG. 2 shows a top plan view of the testing kit  10  with the resealable opening  14  of the plastic bag  13  in the open position. FIG. 3 shows a sectional view of the testing kit  10  taken along line A-A of FIG. 1. The testing kit  10  provides a quick, inexpensive and reliable way to test for the presence of microorganisms  110  (FIG. 4), which may or may not be present in an aqueous solution to be tested  20 , or a suspension to be tested  100 , as fully described below.  
         [0020]    A medium  18  is disposed in the bag interior  17  of the container  12 . The medium  18 , when mixed with an aqueous solution and/or suspension containing microorganisms  110  promotes and fosters the growth of microorganisms  110 , and is nutrient rich to serve this purpose. Currently there are hundreds of different mediums  18  on the market, and they may be in the form of pastes, pellets, powders, liquids, and combinations thereof, and are well known to those skilled in the art.  
         [0021]    As shown in FIG. 4, the aqueous solution to be tested  20 , which may or may not harbor microorganisms  110 , is loaded into the plastic bag  13 .  
         [0022]    In a second embodiment, a suspension to be tested  100 , which may or may not harbor microorganisms  110 , is loaded into the plastic bag  13 , as seen in FIG. 4. In this second embodiment, a suspension to be tested  100  replaces the aqueous solution to be tested  20 . The suspension to be tested  100  may be embodied to comprise, for example, sterile water mixed with soil, milk, or a piece of food like hamburger or chicken. The types of possible suspensions is well known to those skilled in the art.  
         [0023]    In a third embodiment, both an aqueous solution to be tested  20 , and a suspension to be tested  100 , are loaded into the plastic bag  13  simultaneously.  
         [0024]    Thus, the testing kit  10  has the versatility to test for the presence of microorganisms  110  in an aqueous solution to be tested  20 , a suspension to be tested  100 , or both at the same time, all within the same plastic bag  13 . The manner of testing is more fully described below.  
         [0025]    In an embodiment of the testing kit  10 , the medium  18  and aqueous solution to be tested  20 , or suspension to be tested  100 , or both, are placed in the plastic bag  13  and kept at around 37 Degrees Centigrade for 24 hours. This allows any microorganisms  110  in the aqueous solution to be tested  20 , or suspension to be tested  100 , to feed on the medium  18  and multiply many times over, thus making their detection in the at least one reaction chamber  22  more efficient (requiring less time and fewer expensive reagents) (FIG. 1). It is noted that in other embodiments, different temperatures may be equally useful depending on the optimal growth requirements of the microorganisms  110  to be tested.  
         [0026]    The medium  18  may be embodied as comprising a variety of nutrient rich sugars, proteins, and mixtures thereof. A typical growth medium in the plastic bag  13  could contain tryptose, sodium chloride, sorbitol, tryptophan, di-potassium hydrogen phosphate, potassium dihydrogen phosphate, and lauryl sulphate sodium salt plus IPTG (1-Isopropyl-beta-D-1-thiogalactopyranoside). IPTG is an enzyme inducer that induces members of the coliform group of bacteria, if they are present in the aqueous solution to be tested  20 , or suspension to be tested  100 , to excrete enzymes. As described below, the kit  10  can detect the presence of these enzymes.  
         [0027]    Additional examples of mediums  18  for promoting the growth of salmonella and listeria are described in U.S. Pat. No. 5,145,786 to Bailey et al., which is incorporated herein by reference.  
         [0028]    Shown in FIG. 4 is the aqueous solution to be tested  20  mixing with the medium  18 . Of course, as described above, a suspension to be tested  100  could replace the aqueous solution to be tested  20 , as indicated in FIG. 4, or both an aqueous solution to be tested  20  and suspension to be tested  100  may be loaded into the bag  13 .  
         [0029]    [0029]FIG. 1 also shows at least one reaction chamber  22  located in the bag interior  17 . While the present testing kit  10  will work with at least one reaction chamber  22 , it may be embodied with a plurality of reaction chambers  23  as shown in FIGS. 1, 2,  4 . This allows the sequential testing for different microorganisms  110  all within the same plastic bag  13 , without cross contamination between the reaction chambers  23 , as fully described below.  
         [0030]    The at least one reaction chamber  22  may be embodied and constructed of a resilient material. That is, it can be deformed when pressure is applied to it, and then return to its predeformed state when the pressure is relieved. The resilient material may be plastic, but could be constructed of other materials with similar properties, such materials known to those skilled in the art. The at least one reaction chamber  22  has a reactive agent  24  contained therein, the reactive agent  24  is for detecting the presence of microorganisms  110 . The reactive agent  24  may be a plurality of different chromogens or chromogenic substrates. That is, materials that change color when brought into contact with or mixed with, microorganisms  110 , enzymes, microorganism  110  excretions, and so forth. Such reactive agents  24  being well known to those skilled in the art.  
         [0031]    For example, in FIG. 5, the reactive agent  24  in the rupturable membrane reaction chamber  30  may be embodied to contain a chemical such 5-Bromo-4-chloro-03-indoxyl-beta-D-galactopyranoside (“BCIG”). If any coliform bacteria grows in the aqueous solution to be tested  20 , it would excrete the enzyme Beta-D-galactosidase. When this mixes with BCIG in the reaction chamber  30  the BCIG is cleaved by the enzyme, and when so cleaved, a blue-green coloration develops in the aqueous solution to be tested  20 . In this example, BCIG determines the presence of members of the coliform group of bacteria.  
         [0032]    As shown in FIGS. 1 and 2, a plurality of reaction chambers  23  are provided for in the testing kit  10 . The reason for a plurality of reaction chambers  23 , is that it is often desirous to test for many different microorganisms  110  in an aqueous solution to be tested  20 , or a suspension to be tested  100 , and this calls for a different reactive agent  24  for detecting the presence of each different microorganism  110 . Thus, while the reactive agent  24  BCIG determines the presence of the coliform group of bacteria, distinct reactive agents  24  loaded into each of the plurality of reaction chambers  23  could test for distinct microorganisms  110 .  
         [0033]    For example, in FIG. 1, the first of the plurality of reaction chambers  23   a  could test for coliform bacteria as described above. Next, the second of the plurality of reaction chambers  23   b  could test to determine if any of the coliform bacteria were members of the  E. coli  group of bacteria. To do this, in the second of a plurality of reaction chambers  23   b  the chromogenic reactive agent  24  may be embodied as X-Glucuro CHA salt. This chromogen  24  can be cleaved by a second enzyme known as Beta-D-glucuronidase. This enzyme is excreted by any  E. coli  bacteria that may be present in the aqueous solution to be tested  20 . In this scenario, a blue color will develop, when the aqueous solution to be tested  20  is drawn into the reaction chamber  23   b , confirming the presence of  E. coli  bacteria by the presence of first a blue-green color in the first of the plurality of reaction chambers  23   a , followed by a blue color in the second of the plurality of reaction chambers  23   b.    
         [0034]    Additionally, a third test, for example, to determine the presence of salmonella may be conducted in the third of the plurality of reaction chambers  23   c . Contained in this third of a plurality of reaction chambers  23   c  is an embodiment of reactive agent  24  known to react with salmonella.  
         [0035]    In yet another embodiment of the testing kit, the presence of listeria may be detected. In such an embodiment, one of the plurality of reaction chambers  23  may be loaded with reactive agents such as the ones described in U.S. Pat. No. 6,068,988 to Schabert et al. Listeria monocytogenes secrete phosphatidylinositol-specific phospholipase C (1-phosphatidyl-D-myo-inositol inositolphospho-hydrolase or “PI-PLC”). Cleavage of this reactive agent by bacterial PI-PLC results in mainly the formation of inositol 1,2-cyclic phosphate and 5-bromo-4-chloro-3-indoxyl which, after dimersiation, may subsequently be oxidized by atmospheric oxygen or another oxidant. The result may be a deep blue indigo color.  
         [0036]    In other embodiments of the present testing kit  10 , the presence of fecal streptococcus, enterococcus, iron and sulphur bacteria, holerae, and vibro cholerae may be tested for in water and/or wastewater. These and other microorganisms  110  may be present in natural suspensions such as milk, to be tested  100 . They may also be present in or on solid food products such as meat, chicken, fish, or plant products that, when placed in water, form an aqueous suspension to be tested  100 . The testing kit  10  can be used to analyze these types of suspensions to be tested  100 .  
         [0037]    Similarly, the presence of specific microorganisms  110  and groups of microorganisms  110 , for example coliforms, may be tested for on solids such as soils and or any other solid material that when placed in water will form an aqueous solution to be tested  20  and suspension to be tested  100  mixture.  
         [0038]    Again, the reactive agent and medium for each test are well known to those skilled in the art. Similarly, the testing kit  10  may be used to test for the presence of fungi, yeasts, yersinia, campilobacter, staphylococcus aureus, bacillus cereus and vibrio in foods and/or beverages.  
         [0039]    The reactive agents or chromogenic substrates  24  are typically the most expensive component of the testing kit  10 . Thus, by providing the at least one reaction chamber  22  to have a volume only capable of holding about a couple of milliliters of fluid, only a small amount of chromogenic substrate  24  need be used in the at least one reaction chamber  22 , thus allowing the testing kit  10  to be produced inexpensively. Similarly, since the number of microorganisms  110  in the aqueous solution to be tested  20  have been grown to a high concentration prior to activating the plurality of reaction chambers  23 , the reaction time for the calorimetric reactions is rapid and sequential reactions can be carried out in a relatively short period of time.  
         [0040]    The at least one reaction chamber  22  may be embodied in a plurality of shapes. For example, the balloon shape as shown in FIG. 1, or it may be embodied and shaped as an elongated capsule, or other shapes known to those skilled in the art. These examples of shapes for the reaction chamber  22  are only examples and are not intended to limit the scope of the testing kit  10  in any manner.  
         [0041]    As previously described, the testing kit  10  may be used to test an aqueous solution to be tested  20  and a suspension to be tested  100 , as shown in FIG. 4. The aqueous solution to be tested  20 , or suspension to be tested  100 , might originate from any of a variety of sources. For example, they may come from a faucet, a well, a stream, a lake, collected rain water, bottled water, water coolers in homes and/or offices, restaurants, and any other source where water is consumed. Also, the testing kit  10  allows the testing of recreational waters found at swimming holes, swimming pools, and beaches. Microorganisms  110  in recreational waters have been known to sicken people, and the testing kit  10  can be used to test the water before anyone enters the water. Waste water may also be tested as described above.  
         [0042]    The reaction chamber  22  may be embodied in a plurality of shapes, for example FIGS.  5 - 7  show a rupturable membrane reaction chamber  30  embodiment, and FIGS.  8 - 9 A show the resealable aperture reaction chamber  60  embodiment. The testing kit  10  may also be embodied wherein the reaction chamber  22  is fixed to, or unattached to the bag interior  17  of the plastic bag  13 .  
         [0043]    Turning to FIGS.  5 - 7 , the embodiment shown therein has the rupturable membrane reaction chamber  30 . The rupturable membrane reaction chamber  30  has a wall  32 , the wall  32  has a balloon shape but may be embodied to have other shapes. The wall  32  of the rupturable membrane reaction chamber  30  defines an airspace  34 , and extending from the wall  32  is passage tube  36 . The passage tube  36  defines passageway  38 . Extending across the passageway  38  is a rupturable membrane  40 . The airspace  34  of the rupturable membrane reaction chamber  30  may be filled with air  35  or any other suitable fluid medium, preferable gas, well known to those skilled in the art.  
         [0044]    Also contained within the rupturable membrane reaction chamber  30  is the reactive agent  24 , which as discussed above reacts when brought into contact with microorganisms  110 . Prior to use, the rupturable membrane reaction chamber  30  is loaded with reactive agent  24 , and the rupturable membrane  40  positioned across the passageway  38 . Such procedures to accomplish this known to well known to those skilled in the art.  
         [0045]    In use of the testing kit  10  having a rupturable membrane reaction chamber  30 , the resealable opening  14  of the plastic bag  13  is opened, and the aqueous solution to be tested  20 , or suspension to be tested  100 , or both, are added to the plastic bag  13 , and the resealable opening  14  is closed or sealed. Medium  18  is mixed with the aqueous solution to be tested  20 , or suspension to be tested  100 , and the microorganisms  110  in the aqueous solution to be tested  20  to feed upon the medium  18 . A microbial culture in the aqueous solution to be tested  20 , or suspension to be tested  100 , is grown for typically 24 hours at 37° Centigrade. In other embodiments, different temperatures may be employed for growth.  
         [0046]    After this time, the rupturable membrane reaction chamber  30  is crushed, as shown in FIG. 6 by an external pressure, depicted by arrows  50 , applied to the wall  32 . This causes the rupturable membrane  38  to burst. The source of the external pressure, depicted by arrows  50 , would typically be the user&#39;s fingers. As shown in FIG. 6, the air  35  in the air space  34  would become pressurized, such that it would cause the rupturable membrane  40  to burst, and the air  35  would exit the rupturable membrane reaction chamber  30  through the passageway  38  in the passage tube  36 . This flow of exiting air  35  is depicted by arrow  52  in FIG. 6.  
         [0047]    Upon releasing the external pressure, depicted by arrows  50 , the rupturable membrane reaction chamber  30 , because it is resilient, returns to its pre crushed state, and in doing so creates a vacuum or negative pressure therein. When this occurs, the aqueous solution to be tested  20 , or suspension to be tested, or combination thereof, is drawn into the rupturable membrane reaction chamber  30 , indicated by arrows  54  in FIG. 7. The reactive agent  24  then mixes with the aqueous solution to be tested  20 , and/or suspension to be tested. This makes a test mixture  56 , which may or may not result in a chromogenic change in the rupturable membrane reaction chamber  30 , because such a change depends on the presence of predetermined microorganisms  110  in the aqueous solution to be tested  20 , and/or suspension to be tested  100 . If a chromogenic or color change occurs in the test mixture  56 , the user can assess which microorganism  110  is present depending on which reactive agent  24  is in the rupturable membrane reaction chamber  30 , and any colormetric changes that occur.  
         [0048]    The rupturable membrane  38  can be constructed of sufficiently small diameter, such that when the rupturable membrane reaction chamber  30  is filled with the aqueous solution to be tested  20 , and/or suspension to be tested  100 , no leakage occurs from the rupturable membrane reaction chamber  30 . This prevents the test mixture  56  from contaminating the remaining aqueous solution to be tested  20 , or suspension to be tested  100 , in the plastic bag  13 . Thus each of the rupturable membrane reaction chambers  30  can be loaded with a different, distinct reactive agent  24 , to conduct numerous tests on the aqueous solution to be tested  20 , and/or suspension to be tested, without significant cross contamination between the rupturable membrane reaction chambers  30 .  
         [0049]    Another embodiment of the reaction chamber  22  is shown in FIGS.  8 - 10 , which illustrates the resealable aperture reaction chamber  70  embodiment of the testing kit  10 . The resealable aperture reaction chamber  70  has a wall  72  having a balloon shape, however, the wall  72  may be embodied to have a capsule shape, a spherical shape or any other shape known to those skilled in the art. The wall  72  of the resealable aperture reaction chamber  70  defines therein an airspace  74  filled with air  35  or any other suitable gas or fluid medium known to those skilled in the art. Extending from the wall  72  is passage tube  76 . The passage tube  76  defines a passageway  78  therein. Positioned in the passageway  78  and extending fully across said passageway  78  is the resealable aperture  80 , which is made of a resilient material such as resilient plastic, such that after it is deformed, it is able to automatically returns to its predeformed state.  
         [0050]    Also contained within the resealable aperture reaction chamber  70  is the reactive agent  24  that reacts with microorganisms  110 . Prior to use, the resealable aperture reaction chamber  70  is loaded with the reactive agent  24 , and the resealable aperture  80  is closed.  
         [0051]    To use the resealable aperture reaction chamber  70 , pressure, indicated by arrows  50  in FIG. 9, is applied to the wall  72 . Again, this pressure, indicated by arrows  50 , is typically applied by the user&#39;s fingers. As the pressure, indicated by arrows  50 , increases the resealable aperture  80  expands and opens, and air  35  is forced out of the airspace  74 . This air  35  proceeds through the resealable aperture  80  and the passageway  78  in the passage tube  76 . Arrows  52  in FIG. 9 indicated the air  35  exiting the resealable aperture reaction chamber  70 .  
         [0052]    In FIG. 9A, the pressure, indicated by arrows  50 , is relieved, thus not shown in FIG. 9A, and the resealable membrane reaction chamber  70  returns to its prior balloon shape, and in doing so draws the aqueous solution to be tested  20 , or suspension to be tested  100 , into the resealable aperture reaction chamber  70  indicated by arrows  54 . This occurs due to the vacuum or negative pressure generated as the resealable membrane reaction chamber  70  returns to predeformed state as shown in FIG. 8. The resealable aperture  80  closes, thus preventing the test mixture  56  contained in the resealable aperture reaction chamber  70  from leaking, and cross-contaminating any of the other plurality of reaction chambers  23 .  
         [0053]    Just as with the rupturable membrane reaction chamber  30 , chromogenic changes in the test mixture  56  in the resealable aperture reaction chamber  70  may be analyzed for the detection of microorganisms  110 .  
         [0054]    The testing kit  10  may also be embodied to have a plurality of both rupturable membrane reaction chambers  30  and resealable aperture reaction chambers  70 , in the plastic bag  13  at the same time. This is due to the fact that there is no cross contamination between the rupturable membrane reaction chambers  30  and the resealable aperture reaction chambers  70 .  
         [0055]    Another embodiment of the present invention provides for a sequential narrowing process to be used for testing the presence of a specific microorganism, for example a pathogenic variant of the coliform group of bacteria. In this embodiment, separation of specific enzymatic, and biochemical reactions for positive identification of a given species is possible, for example,  E. coli  identified by the presence of both galactosidase and glucoronidase enzymes, or the positive identification of a specific strain of bacteria or pathovar, such as  E. coli  0157.  
         [0056]    For example, a narrowing process may be used to detect  E. coli  bacteria. First, the plastic bag  13  is loaded with three reaction chambers  22  having different reactive agents  24  contained therein, either rupturable membrane reaction chambers  30  or resealable aperture reaction chambers  70 , may be used. The plastic bag  13  is also loaded with the medium  18  and the suspect aqueous solution to be tested  20 .  
         [0057]    The first of the three reaction chambers  22  is crushed, and if there is no reaction, then the specific microorganism  110  being sought is not present. However, if upon crushing there is a reaction in the reaction chamber  22 , then it is known that the generic coliform group of bacteria is present, and the testing continues.  
         [0058]    The type of bacteria is further narrowed by crushing the next reaction chamber  22 . If there is no reaction, the specific type of bacteria being testing for is not present. If a reaction does occur, that narrows the range of the coliform group of bacteria present, and the testing continues.  
         [0059]    The last of the reaction chambers  22  is crushed, and if there is no reaction, this indicates that a pathogenic form of coliform bacteria is not present. If, however, there is a reaction, a pathogenic member of the coliform group of bacteria has been detected, such as  E. coli,  and the user is placed on alert, as this form of the bacteria may be deadly if consumed. The reactive agents  24  used in such sequential testing as described herein, being well known to those skilled in the art.  
         [0060]    The testing kit  10  may also have a kill pod  11 , as shown in FIG. 1, having a sufficient dose of chemicals to kill most, if not all, microorganisms  110 . The kill pod  11  may be embodied as a crushable plastic capsule loaded with chemicals and other materials that destroy microorganisms  110 . The user need only manually crush the kill  11  pod to release the chemicals at the end of the testing to destroy the microorganisms  110  in the aqueous solution to be tested  20 , or suspension to be tested  100 . This step renders the kit  10  safe for disposal following its use to detect potentially pathogenic microorganisms.  
         [0061]    The testing kit  10  also provides for a methodology to use the kit. The method entails multiple steps, for example, the first step requires providing a container  12  which may be embodied as a plastic bag  13 . Second, opening a resealable opening  14  of the container. Third, depositing a medium  18  in the plastic bag  13 . Fourth, placing at least one reaction chamber  22  having a reactive agent  24  contained therein in the plastic bag  13 . This reaction chamber  22  may be a rupturable membrane reaction chamber or a resealable aperture reaction chamber. Fifth, adding the aqueous solution to be tested  20 , or suspension to be tested, to the plastic bag  13 , or both. Sixth, closing the resealable opening  14  in the plastic bag  13 . Seventh, sealing the plastic bag  13 . Eighth, mixing the medium  18  with the aqueous solution to be tested  20 , or suspension to be tested. Ninth, rupturing the rupturable membrane reaction chamber  30  or opening the resealable aperture reaction chamber  70 , or both, depending on which embodiment is being utilized, so that the aqueous solution to be tested  20 , or suspension to be tested, in the plastic bag  13  mixes with the reactive agent  24 . Tenth, examining the test mixture  56  for chromogenic changes. And finally, crushing the kill pod  11  to destroy any the microorganisms  110  present in order to render the kit safe for disposal. All of these steps do not have to be done to utilize the present invention, for example, the use of a kill pod may be unnecessary in all applications.  
         [0062]    It is noted that the embodiments of testing kit  10  are in no way limited to the detection of deleterious microorganism  110 . The present testing kit  10  may be embodied to detect the presence of beneficial microorganisms  110  as well. Beneficial microorganisms  110  may be defined as microorganisms  110  that are beneficial to humans and animals, for example, yeasts for breads and beverages, bacteria for cheeses, bacteria in digestive tracts, etc. The testing kit  10  may be embodied to have the requisite reactive agent  24  and medium  18  to test for the presence of such beneficial microorganisms  110  without departing from the matter disclosed herein and the principles of the testing kit  10 . Such matching of the reactive agent  24  to the microorganism  110  to be tested, being well known to those skilled in the art.  
         [0063]    Thus, the present testing kit  10  provides an inexpensive, easy use, reliable, and quick way to test for a plurality of microorganisms  110 .  
         [0064]    It is understood that, while the invention has been described in detail herein, the invention can be embodied otherwise without from the principles thereof. All of these other embodiments are meant to come within the scope of the present testing kit and methodology as defined in the claims.