WO2016126551A1

WO2016126551A1 - Fluid testing devices and methods of use thereof

Info

Publication number: WO2016126551A1
Application number: PCT/US2016/015709
Authority: WO
Inventors: David Edward Townsend; Shawn Edward DUBOIS
Original assignee: Idexx Laboratories, Inc.
Priority date: 2015-02-05
Filing date: 2016-01-29
Publication date: 2016-08-11

Abstract

The present invention relates to a fluid sample testing device comprising a test strip. One or more separate sample receiving members are attached to the test strip. The sample receiving members include a medium capable of sustaining growth of microorganisms. The sample receiving members are configured to permit a fluid sample to contact the medium when the test strip is contacted with the fluid sample. The fluid testing device also includes a container configured to receive and isolate the test strip. Also disclosed are methods of testing a fluid sample using the device.

Description

FLUID TESTING DEVICES AND METHODS OF USE THEREOF

[0001] This application claims priority benefit of U.S. Provisional Application Serial

No. 62/112,531, filed February 5, 2015, which is hereby incorporated by reference in its entirety.

FIELD OF THE INVENTION [0002] The present invention relates to a fluid sample testing device and a method for testing a fluid sample. More specifically, the present invention relates to devices and methods for detecting the presence of microorganisms in a fluid sample.

BACKGROUND OF THE INVENTION

[0003] Many industries need to detect presence of microorganisms in a fluid sample and to determine possible remediation or treatment solutions. By way of example, such detection may be required for determining the presence of pathogens related to certain infections. Detection also may be utilized for the purposes of water treatment and remediation.

[0004] Specifically, bacterial urinary tract infections are common human and veterinary diseases. Bacteriologic testing is commonly performed on patients experiencing symptoms consistent with urinary tract infections. Microorganisms isolated from patients (human and veterinary) are tested to determine the identity of the pathogens and their susceptibility to antibiotics. The clinical effectiveness of antimicrobial chemotherapy for bacterial urinary tract infections requires the correct identification of the causing pathogens and the selection of an appropriate antibiotic treatment regime to eradicate the disease- causing bacteria. Prior methods for determining the identity of pathogens and treatment options involved isolating the suspect pathogens and incubating the specimen on a culture medium to obtain bacterial growth. The suspect pathogens were then subcultured and their identity and antimicrobial susceptibility were determined using a series of biochemical tests and standards antimicrobial susceptibility tests.

[0005] Due to the time intensive nature of the antimicrobial susceptibility tests, devices and methods that eliminate the need for microbial growth and subsequent testing, and allow for the concurrent determination of presence of a microorganism in a sample and the antimicrobial susceptibility of the microbial organisms have been provided. For example, such devices are presented in U.S. Patent No. 6,984,499 and 7,384,778, which are hereby incorporated herein by reference in their entirety. Prior solutions, however, require specific devices which can render the testing too expensive.

[0006] For example, such testing may be utilized to diagnose and/or determine treatment options for urinary tract infections in the veterinary setting. High costs of the testing may deter some pet owners from proceeding with the test to confirm an infection. Thus, a more cost effective test would encourage owners to complete the testing and improve treatment.

[0007] The present invention is directed to overcoming these and other deficiencies in the art.

SUMMARY OF THE INVENTION [0008] One aspect of the present invention relates to a fluid sample testing device comprising a test strip. One or more separate sample receiving members are attached to the test strip. The sample receiving members include a medium capable of sustaining growth of microorganisms. The sample receiving members are configured to permit a fluid sample to contact the medium when the test strip is contacted with the fluid sample. The fluid testing device also includes a container configured to receive and isolate the test strip.

[0009] Another aspect of the present invention relates to a method for testing a fluid sample comprising providing a fluid sample testing kit. The fluid sample testing kit includes a test strip with one or more separate sample receiving members attached to the test strip. The sample receiving members include a medium capable of sustaining growth of microorganisms. The sample receiving members are configured to permit a fluid sample to contact the medium when the test strip is contacted with the fluid sample. The fluid sample testing kit also includes a container configured to receive and isolate the test strip. A fluid sample is placed in the container. The test strip is inserted into the container, whereby the test strip is not fully immersed in the fluid sample. The fluid sample is caused to contact the one or more receiving members. The test strip is analyzed to determine if the microorganism is present in the fluid sample.

[0010] A further aspect of the present invention relates to a fluid sample testing device comprising a test strip. One or more separate sample receiving members are attached to the test strip. The sample receiving members include medium capable of sustaining growth of microorganisms. The sample receiving members are configured to permit a fluid sample to contact the medium when the test strip is contacted with the fluid sample. A negative control member is attached to the test strip. The negative control member includes the medium and is sealed to prevent contact between the medium and the fluid sample contacting the test strip.

[0011] Yet another aspect of the present invention relates to a method for testing a fluid sample comprising providing a fluid sample testing kit. The fluid sample testing kit includes a test strip with one or more separate sample receiving members attached to the test strip. The sample receiving members include a medium capable of sustaining growth of microorganisms. The sample receiving members are configured to permit a fluid sample to contact the medium when the test strip is contacted with the fluid sample. The fluid sample is caused to contact the one or more receiving members. The test strip is analyzed to determine if the microorganism is present in the fluid sample.

[0012] The fluid testing devices provide an easy to use and economically produced devices for testing a fluid sample to determine the presence of microorganisms in the fluid. The fluid testing devices may also be utilized to simultaneously determine one or more treatment options in the event the microorganisms are present in the fluid. The efficacy of various potential treatment options may further be determined at the same time to streamline the treatment process. Thus, the fluid sample testing devices provide significant cost-savings over available testing methods. Specifically, the fluid sample testing devices may be utilized to detect, and determine treatment options for, urinary tract infections in the veterinary setting. The ease of use and increased cost savings will encourage more frequent testing for such infections, which may have gone undiagnosed due to prohibitive costs. The fluid sample testing devices of the present invention further provide the necessary devices for testing in a single package. The fluid sample testing devices are compact and easily storable.

BRIEF DESCRIPTION OF THE DRAWINGS

[0013] FIG. 1 is a perspective view of an embodiment of a fluid sample testing device of the present invention.

[0014] FIG. 2 is a bottom view of the embodiment of the fluid sample testing device of FIG. 1.

[0015] FIG. 3 is a top view of the embodiment of the fluid sample testing device of

FIG. 1.

[0016] FIG. 4 is a side view of the embodiment of the fluid sample testing device of

FIG. 1. [0017] FIG. 5 is an end view of the embodiment of the fluid sample testing device of

FIG. 1.

[0018] FIG. 6 is a perspective view of another embodiment of a fluid sample testing device of the present invention.

[0019] FIG. 7 is a perspective view of another embodiment of a fluid testing device of the present invention.

[0020] FIG. 8 is an exploded, perspective view of a fluid sample testing kit including the fluid sample testing device of FIG. 1 and a testing tube.

[0021] FIG. 9 is a perspective view of the fluid testing kit of FIG. 8 with the fluid sample testing device inserted into the testing tube in accordance with a fluid testing method of the present invention.

[0022] FIG. 10 is a perspective view of the fluid testing kit of FIG. 8 with the fluid sample testing device inserted into the testing tube which is inverted.

[0023] FIG. 11 A illustrates a portion of a fluid sample testing device including a receiving well and a negative control well prior to testing.

[0024] FIGS. 1 IB and 11C illustrate the portion of the fluid sample testing device of

FIG. 11A after testing.

[0025] FIG. 12 illustrates testing results for determining the presence of

microorganisms and one or more treatment options in accordance with the present invention.

DETAILED DESCRIPTION OF THE INVENTION [0026] The present invention relates to a fluid sample testing device and a method for testing a fluid sample using the device.

[0027] One aspect of the present invention relates to a fluid sample testing device comprising a test strip. One or more separate sample receiving members are attached to the test strip. The sample receiving members include a medium capable of sustaining growth of microorganisms. The sample receiving members are configured to permit a fluid sample to contact the medium when the test strip is contacted with the fluid sample. The fluid testing device also includes a container configured to receive and isolate the test strip.

[0028] FIGS. 1-5 show perspective, bottom, top, side, and end views, respectively, of one embodiment of a fluid sample testing device 10 of the present invention. Fluid testing device 10 may be utilized to test fluids, such as, by way of example only, blood and blood products, plasma, saliva, sweat, semen, vaginal fluid, cerebral spinal fluid, water, or urine, or cell-free samples (e.g., cell extracts, fractions of cell extracts, or cell lysates), histological or fixed samples, tissues or tissue extracts, environmental samples, or whole cells, for the presence of microorganisms or pathogens.

[0029] Fluid sample testing device 10 includes a test strip 12, one or more separate sample receiving members 14, an optional negative control member 16, and an optional elongate shaft 18. Test strip 12 can be an elongate, rectangle configured to support one or more separate sample receiving members 14 and optional negative control member 16, which are attached to test strip 12.

[0030] In one embodiment, test strip 12 is a plastic blister pack with one or more sample receiving members 14 and negative control member 16 as wells integrally formed as the blister portions in the test strip 12. The wells extend from test strip 12 and may be configured to hold a pre-determined amount of sample therein. The wells include an opening suitable to receive the sample into the receiving well. Other methods of attaching one or more sample receiving members 14 and optional negative control member 16 to test strip 12 may be utilized.

[0031] By way of example, in another embodiment, the receiving members 14 may be absorbent pads attached to the test strip 12, as illustrated in FIG. 6. Absorbent pads are constructed of a material suitable to absorb a liquid sample that comes into contact with the pads. The absorbent pads may be attached to the test strip using an adhesive, by way of example, or may be attached by a friction fit. Four sample receiving members 14 are illustrated in FIGS. 1-4 (receiving wells), and 6 (absorbent pads); however, fluid sample testing device 10 may include more or fewer sample receiving members 14. Sample receiving members 14 may be designed to retain a predetermined amount of fluid when contacted with a fluid sample.

[0032] Referring again to FIGS. 1-5, each of the one or more sample receiving members 14 includes medium 20, which is capable of sustaining growth of microorganisms or pathogens, located therein. Medium 20 may generally include one or more of a carbon source, nitrogen source, amino acids, and various salts for pathogen growth. In certain embodiments, medium 20 may be a selective medium, which selectively promotes the growth of a specific pathogen by an antibacterial/antifungal effect, temperature or pH resistance, or the ability to synthesize a certain metabolite, thereby allowing the pathogen to grow selectively vis-a-vis other competing bacteria or microorganisms. In some embodiments, the pathogen detection reagent may contain compounds such as L-cysteine and Oxyrase

(Oxyrase Inc., Mansfield, Ohio) to accelerate the growth of specific pathogens or resuscitation of injured pathogens by reducing oxygen concentration in the medium 20. In certain embodiments, the medium 20 may he a differential medium, which is formulated to display a color change when the microorganisms growing metabolize a certain ingredient.

[0033] The medium 20 can include any growth medium known to those skilled in the art, including MacConkey's (MAC) media, Mannitol Salt media, Blood Agar, Hektoen enteric (HE) agar, Terrific Broth (TB), Xylose lysine desoxyscholate (XLD), or

Streptococcuseosin methylene blue (EMB). In one embodiment, medium 20 allows for the substantial growth of only a specific target pathogen and for substantially less growth of any other bacteria. The specific pathogen may include, by way of example, skin infection organisms, ear infection organisms, or other organisms that cause a disease or condition of medical interest. In one embodiment, medium 20 allows for the substantial growth of the primary Gram negative and positive uropathogens, which are the primary causative agents of urinary tract infections. The Gram negative and/or Gram positive uropathogens may include Escherichia coli, Kebsiella spp., Proteus spp., Pasteur ella spp., Pseudomonas spp.,

Enter ococcus spp., Staphylococcus spp., Streptococcus spp., Pantoae spp., Enter obacter sp., Serratia spp. , Citrobacter spp. , Acinetobacter spp. , Providencia spp., Morganella morganii, and Salmonella enterica, although medium 20 may allow substantial growth of other uropathogens.

[0034] The presence of the microorganism can be detected using optical detection methods. In some embodiments, optical detection method includes detecting at least one color change.

[0035] The presence of the microorganism can be detected by essentially any desired optical detection method that is currently known or hereafter developed. Such methods include, for example, human visual inspection, luminescence detection, fluorescence detection, microscopy (for example, using a transmitted light microscope or an

epifluorescence microscope, which can be used for visualizing microorganisms tagged with fluorescent dyes), other methods of analog or digital optical imaging (based, for example, on reflection, absorption, transmission, and/or luminance measurements by an imaging device such as, for example, a camera, video equipment, or a scanner), and the like, and

combinations thereof. Preferred methods include human visual inspection, digital optical imaging (more preferably, digital optical imaging using a scanner), and combinations thereof. In one embodiment, medium 20 provides a detectable signal, such as, by way of example only, a color change, when substantial growth of the target pathogen occurs. In one embodiment, each of one or more sample receiving members 14 are receiving wells that further include an opening 22 suitable to permit a fluid sample to contact medium 20 therein when test strip 12 is contacted with a fluid sample. In another embodiment, as shown in FIG. 6, medium 20 is impregnated within the absorbent pad. Absorbent pad may absorb the liquid sample through a surface thereof to contact medium 20 located inside the absorbent pad. Absorbent pad is constructed of an absorbent material suitable to allow the fluid sample to contact the medium through the surface of the absorbent pad, while providing an outer shell to contain the medium therein.

[0036] Referring again to FIGS. 1-5, optional negative control member 16 also includes medium 20 therein. Negative control member 16 further includes a seal 24, such as, by way of example only, a foil wrapper, to prevent a fluid sample from contacting medium 20 therein when test strip 12 is contacted with a fluid sample.

[0037] Another aspect of the present invention relates to a fluid sample testing device wherein one or more, but not all, of the one or more sample receiving members 14 further include an antimicrobial agent. Referring now to FIG. 7, fluid sample testing device 10 is illustrated. Fluid sample testing device 10 is the same in structure and operation as described above, except as illustrated and described herein.

[0038] In this embodiment, fluid sample testing device 10 includes fluid sample receiving members 14(a)- 14(d), although fluid sample testing device 10(2) may include more or fewer fluid sample receiving members in other configurations. Although receiving members 14(a)-14(d) are illustrated as receiving wells, it is to be understood that the receiving members 14(a)-14(d) could be absorbent pads as illustrated in FIG. 6. Fluid sample receiving member 14(a) includes medium 20 and serves as a positive control for determining the presence of microorganisms, as described further below. Fluid sample receiving members 14(b)- 14(d) further each include a predetermined amount of a different

antimicrobial agents 25(a)-25(c) located within medium 20. Antimicrobial agents include, but are not limited to, antibacterial agents, antiviral agents, antifungal agents, or anti-parasitic agents.

[0039] Antibacterial agents include beta-lactam compounds such as penicillin, methicillin, nafcillin, oxacillin, cloxacillin, dicloxacillin, ampicillin, ticarcillin, amoxicillin, carbenicillin, and piperacillin; cephalosporins and cephamycins such as cefadroxil, cefazolin, cephalexin, cephalothin, cephapirin, cephradine, cefaclor, cefamandole, cefonicid, cefuroxime, cefprozil, loracarbef, ceforanide, cefoxitin, cefmetazole, cefotetan, cefoperazone, cefotaxime, ceftazidine, ceftizoxine, ceftriaxone, cefixime, cefpodoxime, proxetil, cefdinir, cefditoren, pivoxil, ceftibuten, moxalactam, and cefepime; other beta-lactam drugs such as aztreonam, clavulanic acid, sulbactam, tazobactam, ertapenem, imipenem, and meropenem; other cell wall membrane active agents such as vancomycin, teicoplanin, daptomycin, fosfomycin, bacitracin, and cycloserine; tetracyclines such as tetracycline, chlortetracycline, oxytetracycline, demeclocycline, methacycline, doxycycline, minocycline, and tigecycline; macrolides such as erythromycin, clarithromycin, azithromycin, and telithromycin;

aminoglycosides such as streptomycin, neomycin, kanamycin, amikacin, gentamicin, tobramycin, sisomicin, and netilmicin; sulfonamides such as sulfacytine, sulfisoxazole, silfamethizole, sulfadiazine, sulfamethoxazole, sulfapyridine, and sulfadoxine;

fluoroquinolones such as ciprofloxacin, gatifloxacin, gemifloxacin, levofloxacin,

lomefloxacin, moxifloxacin, norfloxacin, and ofloxacin; antimycobacteria drugs such as isoniazid, rifampin, rifabutin, rifapentine, pyrazinamide, ethambutol, ethionamide, capreomycin, clofazimine, and dapsone; and miscellaneous antimicrobials such as

colistimethate sodium, methenamine hippurate, methenamine mandelate, metronidazole, mupirocin, nitrofurantoin, polymyxin B, clindamycin, choramphenicol, quinupristin- dalfopristin, linezolid, spectrinomycin, trimethoprim, pyrimethamine, and trimethoprim- sulfamethoxazole. The antibacterial agents also include benzoic acid, sodium benzoate, isopropyl p-hydroxybenzoate, isobutyl p-hydroxybenzoate, ethyl p-hydroxybenzoate, methyl p-hydroxybenzoate, butyl p-hydroxybenzoate, propyl p-hydroxybenzoate, sodium sulfite, sodium hyposulfite, potassium pyrosulfite, sorbic acid, potassium sorbate, sodium

dehydroacetate, thujaplicin, udo extract, storax extract, wild tansy extract, milt protein extract, zymolytic Job's tears extract, and the like. Antibacterial agents also can include retapamulin, silver sulfadiazine, benzol peroxide, hydrogen peroxide, clindamycin phosphate, minocycline, azelaic acid, sodium sulfacetamide, sodium sulfacetamide sulfur, neramexane, carbapenems, monobactams, chloramphenicol, glycylcylines, ketolide (telithromycin), spectinomycin, lincosamides, colistin, and novobiocin.

[0040] Antifungal agents include a polyene, an azole, an allylamine (for instance, naftifine or terbinafine) a benzyl amine (for instance, butenafine), or other antifungal agents (for instance, amorolfine). Examples of polylenes include, e.g., nystatin and Amphotericin B. Examples of azoles include miconazole, clotrimazole, ketoconazole, oxiconazole,

eberconazole, econazole, sulconazole and sertaconazle bifonazole, butoconazole,

fenticonazole, isoconazole, omoconazole and tioconazole, or pharmaceutically acceptable salts thereof. Still other antifungal agents include, e.g., ciclopirox or selenium sulfide.

Additional antifungals include agents that block NA synthesis including, e.g., flucytosine, and those that disrupt microtubule function including, e.g., griseofulvin. Suitable antifungals can include one of candicidin, filipin, hamycin, natamycin, and rimocidin. Triazoles, including albaconazole, fluconazole, isavuconazole, itraconazole, posaconazole,

ravuconazole, terconazole, and voriconazole are also suitable antifungal active agents. Also suitable are, thiazoles including, e.g., abafungin. In addition, echinocandins, including anidulafungin, caspofungin, and micafungin, are suitable antifungals. Also suitable are griseofulvin, benzoic acid, ciclopirox, haloprogin, polygodial, tolnaftate, undecylenic acid, and Crystal violet.

[0041] Anti-parasitic agents include permethrin, ivermectin, pyrethrins, lindane, malathion, benzyl benzoate, thiabendazole or diiodohydroxyquinoline (iodoquinol). Non- limiting examples of anti-parasite agents include antimalaria drugs such as chloroquine, amodiaquine, quinine, quinidine, mefloquine, primaquine, sulfadoxine-pyrimethamine, atovaquone-proguanil, chlorproguanil-dapsone, proguanil, doxycycline, halofantrine, lumefantrine, and artemisinins; treatments for amebiasis such as metronidazole, iodoquinol, paromomycin, diloxanide furoate, pentamidine, sodium stibogluconate, emetine, and dehydroemetine; and other anti-parasite agents such as pentamidine, nitazoxanide, suramin, melarsoprol, eflornithine, nifurtimox, clindamycin, albendazole, and tinidazole.

[0042] Antiviral agents include acyclovir, pencyclovir, cidofovir, idoxuridine, stavudine, zidovudine, ribavarin, amantadine, foscarnet, didanosine, acyclovir, ganciclovir, cidofovir, zalcitabine, rimantadine, calacyclovir, famiciclovir, abacavir, didanosine, emtricitabine, lamivudine, stavudine, tenofovir, zalcitabine, zidovudine, zidovudine- lamivudine, TRIZIVIR (zidovudine, lamivudine, abacavir), EPZICOM (aba-cavir- lamivudine), TRUVADA (tenofovir-emtricitabine), efavirenz, nevirapine, and delavirdine, amprenavir, atazanavir, fosamprenavir, indinavir, lopinavir-ritonavir, nelfinavir, ritonavir, saquinavir, and tipranavir.

[0043] In one embodiment, the different antimicrobial agents 25(a)-25(c) may be amoxicillin, clavulanic acid/amoxicillin, or enrofloxacin, although other antimicrobial agents may be utilized.

[0044] A further aspect of the present invention relates to a method of testing a fluid sample comprising providing a fluid sample testing kit. The fluid sample testing kit includes a test strip with one or more separate sample receiving members attached to the test strip. The sample receiving members include a medium capable of sustaining growth of microorganisms. The sample receiving members are configured to permit a fluid sample to contact the medium when the test strip is contacted with the fluid sample. The fluid sample testing kit also includes a container configured to receive and isolate the test strip. A fluid sample is placed in the container. The test strip is inserted into the container, whereby the test strip is not fully immersed in the fluid sample. The fluid sample is caused to contact the one or more receiving members. The test strip is analyzed to determine if the microorganism is present in the fluid sample.

[0045] Turning now to FIG. 8, a fluid sample testing kit 100 including fluid sample testing device 10 is illustrated. Fluid sample testing kit 100 further includes an container, such as elongate tube 26 configured to receive a fluid sample (S) and fluid sample testing device 10. Although the container is described as an elongate test tube, it is to be understood that other containers configured to receive and isolate the test strip may be utilized, such as petri dishes, flasks, or centrifuge tubes, by way of example only. The container isolates the test strip by reducing the likelihood of evaporation (drying out) or contamination of the test strip after insertion into the container.

[0046] In this example, elongate tube 26 includes a hollow tube extending between open end 28 and closed end 30, which is located opposite open end 28. Optional elongate shaft 18 is configured to maintain a space between closed end 30 of elongate tube 26 and test strip 12 as illustrated in FIG. 9, thereby preventing test strip 12 from being immersed in the fluid sample (S) upon insertion into elongate tube 26. Cap structure 32 may be utilized to seal open end 28 of elongate tube 26 to allow for inversion of elongate tube 26 to contact fluid sample (S) with test strip 12 as illustrated in FIG. 10. Fluid testing kit 100 may further include sterile pouch 34 for housing test strip 12 prior to insertion into elongate tube 26. Sterile pouch 34 prevents medium 20 stored in receiving members 14 from interacting with microorganisms prior to testing.

[0047] With reference now to FIGS. 8-10, one embodiment of the method of testing a fluid sample involves placing fluid sample (S) in elongate tube 26. In one embodiment, 0.2 mL of fluid sample (S) is added to elongate tube 26, although other amounts of fluid sample (S) may be utilized. Fluid sample (S), by way of example, may be a urine sample for which the presence of urinary pathogens is to be determined, although the present method may be utilized with other fluids as described above to determine the presence of microorganisms or pathogens in the fluid.

[0048] Next, fluid sample testing device 10 is removed from sterile pouch 34, which is utilized to store fluid sample testing device 10(1), and is inserted into elongate tube 26 though open end 28. Fluid sample testing device 10(1) is inserted into fluid sample testing device 10 with optional elongate shaft 18 positioned toward closed end 30 of elongate tube 26 as shown in FIG. 8, such that test strip 12 is not fully immersed in fluid sample (S), as shown in FIG. 9.

[0049] After insertion of fluid sample testing device 10 into elongate tube 26, cap structure 32 is secured to open end 28 to seal elongate tube 26. Fluid sample (S) is then caused to contact medium 20 in receiving members 14. In one embodiment, receiving members 14 are configured to retain a pre-determined amount of fluid sample (S). Fluid sample (S) may be caused to contact receiving members 14, by way of example, through inversion of elongate tube 26 as shown in FIG. 10, although other methods may be utilized to inoculate medium 20. In one embodiment, elongate tube 26 is inverted at least ten times, although more or fewer inversions may be employed. In one embodiment, after test strip 12 is initially in contact with fluid (S) (for example, by dipping the test strip into the fluid (S), which causes the receiving members 14 to retain a pre-determined amount of fluid (S), the fluid (S) must be separated from test strip 12 to avoid further contact. In one example, the remaining fluid (S) can be removed from elongate tube 26 by various means or can remain in the elongate tube 26 as long as it is no longer in physical contact with sample receiving members 14 of test strip 12. Retaining the remaining liquid in elongate tube 26 can help in maintaining the humidity around test strip 12, thereby preventing test strip 12 from drying out during incubation. In another embodiment, an absorbent material is present in elongate tube 26 to contain the fluid (S) after the initial contact with the sample receiving members 14.

[0050] Next, test strip 12 is allowed to incubate for an appropriate amount of time and at a temperature to cause any viable microorganisms present in fluid sample (S) to grow on or in medium 20 in sample receiving members 14. Elongate tube 26 is placed in an upright position, as show in FIG. 9, during the incubation. In one embodiment, the test strip is incubated for 24 hours at 35 degrees Celsius, although other incubation periods, at other temperatures, may be utilized.

[0051] After the incubation period, test strip 12 is analyzed to determine if the microorganism is present in fluid sample (S). In one embodiment, the analyzing includes determining if the microorganism is present in fluid sample (S) based on a comparison of medium 20 in sample receiving members 14 and medium 20 in negative control well 16. The presence of the microorganism can be detected using optical detection methods as described above. In some embodiments, the optical detection method includes detecting at least one color change. For example, a color change of medium 20 in sample receiving members 14 as compared to medium 20 in negative control well 16, which is protected from contact with fluid sample (S) by seal 24, may be detected. [0052] Another aspect of the present invention relates to a method for testing a fluid sample comprising providing a fluid sample testing kit. The fluid sample testing kit includes a test strip with one or more separate sample receiving members attached to the test strip. The sample receiving members include a medium capable of sustaining growth of microorganisms. The sample receiving members are configured to permit a fluid sample to contact the medium when the test strip is contacted with the fluid sample. The fluid sample is caused to contact the one or more receiving members. The test strip is analyzed to determine if the microorganism is present in the fluid sample.

[0053] In this embodiment, receiving members 14 may be contacted by fluid sample

(S) by inserting test strip 12 into fluid sample (S), which is held in a container, such as by dipping test strip 12 into fluid sample (S) for a period of time. The test strip 12 is then removed from fluid sample (S). In another embodiment, test strip 12 may be exposed to a stream of fluid sample (S) to provide contact between fluid sample (S) and sample receiving members 14. The test strip 12 may then be stored in the absence of fluid sample (S) for incubation and analysis. In one embodiment, test strip 12 can be contacted with fluid sample (S), sealed in a container, and shipped to an off-site facility for analysis. During shipment, fluid sample (S) and medium 20 will incubate, thereby saving time prior to reading the result. A time and date of fluid sample (S) contact are placed on or with the device to inform the testing facility of the incubation time and appropriate time to take a reading.

[0054] Referring now to FIGS. 11 A-l 1C, sample testing results are illustrated. FIG. 11 A shows a portion of fluid sample testing strip 12 prior to testing. As shown in FIG. 1 IB, which illustrates the portion of fluid sample testing strip 12 shown in FIG. 11 A, after the incubation period, medium 20 in receiving member 14 remained the same color as medium 20 in negative control member 16, indicating a negative result for microorganisms, i.e., no growth of microorganisms on medium 20 in receiving member 14. FIG. 11C illustrates two positive results for microorganism growth. As shown in FIG. 11C, medium 20 in receiving member 14 has changed in color as compared to negative control member 16 as a result of microorganism growth. The change in color indicates the presence of the microorganism in fluid sample (S).

[0055] Another aspect of the present invention involves utilizing fluid testing device 10(2) and determining one or more treatment options based on a comparison of medium 20 in sample receiving member 14(1), without an antimicrobial agent, and medium 20 in sample receiving members 14(2)-14(3), which include antimicrobial agents 25(a)-25(c). [0056] Turning now to FIG. 12, sample testing results using fluid sample testing device 10 are illustrated. Fluid sample receiving member 14(a) serves as a positive control. The color change in fluid sample receiving member 14(a), which does not include an antimicrobial agent, as compared to negative control member 16 indicates the presence of a microorganism in fluid sample (S), i.e., the growth of microorganisms. Based on the determined presence of the microorganism, the color change in fluid sample receiving members 14(b) and 14(c) indicates that the microorganisms are resistant to antimicrobial agents 25(a) and 25(b). Further, the lack of color change in fluid sample receiving member 14(c) indicates that the microorganisms are susceptible to antimicrobial agent 25(c), i.e., the presence of antimicrobial agent 25(c) resulted in to microorganism growth in fluid sample receiving member 14(c). Therefore, antimicrobial agent 25(c) is indicated as a treatment option for the microorganisms.

EXAMPLES

[0053] The following examples are intended to illustrate practice of the invention, and are not intended to limit the scope of the claimed invention. Materials and Methods

[0054] Preparation of 2x liquid medium for fluid sample testing: Liquid medium is first prepared at a 2x concentration. 2x concentrated medium #1 was prepared by combining HEPES free acid, 7.00 grams/liter; HEPES sodium salt, 4.00 grams/liter; magnesium sulfate, 0.20 grams/liter; sodium carbonate, 1.00 grams/liter; yeast extract, 5.00 grams/liter; proteose peptone # 3, 10.00 grams/liter; D-(+)-glucose, 5.00 grams/liter; casein peptone, 20.00 grams/liter; resazurin, 0.10 grams/liter; and methylene blue, 0.02 grams/liter. 2x

concentrated medium was filtered through a 0.2 μπι filter assembly.

[0055] Preparation of lx agar medium for fluid sample testing: Briefly, 30 grams/liter of agar was heat sterilized by autoclave for 15 minutes at 121°C. lx medium was prepared by combining 25 ml of 2x liquid medium with 25 ml of the 2x agar solution in one of four solution vessels corresponding to the following conditions: no antibiotic, 0.0240 grams/liter amoxicillin, 0.0500 grams/liter cephalothin, or 0.0045 grams/liter ciprofloxacin. The vessels were then shaken gently to thoroughly combine the medium and agar solutions.

[0056] Preparation of the fluid sample testing device: 80 μΐ of lx agar medium was transferred into each corresponding sample receiving well of the fluid sample testing device. The top well of the fluid sample receiving device was sealed and serves as a negative control. The second, third, fourth, and fifth sample receiving wells correspond to a viability well containing no antibiotics, an amoxicillin well, a cephalothin well, and a ciprofloxacin well, respectively.

[0057] Microorganism detection assay: Samples were analyzed in duplicate. 2.7 ml of sterile saline was first added to a 15 ml conical tube. Next, 300 μΐ of a urine sample was added to the tube, generating a 1 : 10 dilution of the original sample. The fluid sample testing device was then transferred to the 15 ml conical tube. Care was taken to ensure the testing wells were elevated above the diluted sample. The conical tube was then capped and mixed by inversion 10 times in order to inoculate each well. The device was returned to the upright position and incubated for 24 hours at 35°C. Urine samples are cultivated on lx agar medium in the presence or absence of antibiotics. At the termination of the incubation period, the fluid sample testing device is removed from the conical tube and examined for color changes compared to the negative control well. Any change in color relative to the negative control well is considered positive and indicates growth.

Example 1: Detection of Microorganisms in a Urine Sample Using Growth Medium

[0058] The presence or absence of a microorganism in several urine samples was evaluated using a fluid sample testing device containing the growth medium. The ingredients of the growth medium are listed in Table 1 below:

Table 1: GROWTH MEDIUM

MATERIAL Grams /Liter

HEPES, FREE ACID 3.500

HEPES, Na 2.000

MAGNESIUM SULFATE 0.100

SODIUM CARBONATE 0.500

YEAST EXTRACT 2.500

PROTEOSE PEPTONE #3 5.000

D-(+)-GLUCOSE 2.500

PEPTONE, CASEIN 10.000

RESAZURIN 0.050

METHYLENE BLUE 0.010

AGAR 15.000

AMOXICILLIN 0.024

CEPHALOTHIN 0.050

CIPROFLOXACIN 0.0045 [0059] Each urine sample was evaluated in duplicate. Results are shown in Table 2 below. Microorganisms were detected in all of the viability wells.

Table 2: Microorganism Detection Using Growth Medium

Viability

Sample ID Amoxicillin Cephalothin Ciprofloxacin cfu/mL

Cube

3948689 +/+ +/+ +/+ +/+ >1.00E+08

3956260 +/+ +/+ +/+ +/+ >1.00E+08

3961555 +/+ +/+ +/+ +/+ >1.00E+08

3961036 +/+ +/+ +/- +/+ >1.00E+08

4025278 +/+ -/- -/- +/+ >1.00E+08

4038472 +/+ -/- -/- -/- >1.00E+08

4036422 +/+ -/- -/- -/- >1.00E+08

3999964 +/+ +/+ +/+ -/- >1.00E+08

3962703 +/+ -/+ +/+ +/+ >1.00E+08

4038436 +/+ +/+ +/+ +/+ >1.00E+08

4186493 +/+ -/+ +/+ +/+ >1.00E+08

4207753 +/+ +/+ +/+ +/+ >1.00E+08

4172621 +/+ +/+ +/+ +/+ >1.00E+08

4197414 +/+ +/+ +/+ +/+ >1.00E+08

4206193 +/+ +/+ +/+ +/+ >1.00E+08

4245923 +/+ -/- +/+ +/+ >1.00E+08

4177162 +/+ -/- 7+ -/- >1.00E+08

4245478 +/+ -/+ +/+ +/+ >1.00E+08

4565735 +/+ -/- +/+ +/+ >1.00E+08

4720091 +/+ -/- -/- +/+ >1.00E+08

5257642 +/+ +/+ +/+ +/+ 3.15E+08

4953235 +/+ +/+ +/+ -/+ 2.87E+08

4967769 +/+ -/- 7+ +/+ 2.45E+08

2019647 +/+ -/- +/+ -/+ 2.23E+08

R1990413 +/+ -/- +/+ +/+ 1.89E+08

4701873 +/+ * -/- -/- -/+ 1.87E+08

5147767 +/+ -/- 7+ +/+ 1.27E+08

4690018 +/+ * -/- -/- -/- 1.08E+08

5267246 +/+ +/+ +/+ +/+ 9.80E+07

Q7828565 +/+ +/- +/+ +/+ 2.94E+07

3949149 +/+ +/+ -/- -/- 2.56E+07

4600683 +/+ -/- -/- +/+ 2.46E+07

4185692 +/+ -/- +/- +/+ 2.29E+07

R1687851 +/+ +/+ +/+ +/+ 2.02E+07

5108212 +/+ 7- 7- +/+ 1.95E+07 5157656 +/+ -/- -/- -/- 1.71E+07

5066589 +/+ +/+ +/+ +/+ 1.60E+07

5905724 +/+ -/- -/- -/- 9.80E+06

5094144 +/+ -/- -/- +/+ 5.20E+06

4551533 +/+ -/- +/+ -/- 2.54E+06

4221805 +/+ +/+ +/+ +/+ 2.03E+06

6843223 +/+ +/+ +/+ -/- 1.13E+06

4561567 +/+ +/+ +/+ -/- 5.50E+05

4209481 +/+ -/- -/- -/- 2.68E+05

6616922 +/+ +/+ +/+ +/+ 1.12E+05

6062234 +/+ -/- -/- -/- 1.00E+05

6662442 +/+ -/- -/- -/- 3.10E+04

4240524 +/+ +/+ +/+ -/- 3.00E+04

3978122 +/+ +/+ +/+ +/+ 2.15E+04

4194181 +/+ -/- -/- -/- 2.01E+04

4177171 +/+ -/- +/+ -/- 1.60E+04

4558884 +/+ -/- -/- +/+ 9.20E+03

4686839 +/+ -/- -/- -/+ 4.00E+03

4729417 +/+ -/- -/- +/- 3.40E+03

5274699 +/+ -/- -/- -/- 1.70E+03

6067357 +/+ -/- -/- -/- 1.20E+03

* Sample was viewed at approximately six hours and noted to have a positive signal in the viability cube. Appearance of color change compared to uninoculated medium; '+' = Bacterial Growth, '-' = No Bacterial Growth; CFU = Colony Forming Unit.

[0060] In the case of determining the presence or absence of urinary tract infections from a urine sample, it has been found that some samples having >10⁸ cfu/mL gave a positive signal in about six hours.

[0061] Although preferred embodiments have been depicted and described in detail herein, it will be apparent to those skilled in the relevant art that various modifications, additions, substitutions, and the like can be made without departing from the spirit of the invention and these are therefore considered to be within the scope of the invention as defined in the claims which follow.

Claims

WHAT IS CLAIMED:

1. A fluid sample testing device comprising:

a test strip;

one or more separate sample receiving members attached to the test strip, each of said one or more sample receiving members comprising a medium capable of sustaining growth of microorganisms and configured to permit a fluid sample to contact the medium when the test strip is contacted with the fluid sample; and

a container configured to receive and isolate the test strip.

2. The fluid sample testing device of claim 1, wherein the container is selected from the group consisting of petri dishes, flasks, test tubes, and centrifuge tubes.

3. The fluid same testing device of claim 2, wherein the container is a test tube having an oening at one end, being closed at an opposite end. 4. The fluid sample testing device of claim 3, further comprising:

an elongate shaft coupled to the test strip, said elongate shaft configured to maintain space between the closed end of the test tube and the test strip.

5. The fluid sample testing device of claim 3 further comprising:

a cap structure configured to seal the opening of the test tube.

6. The fluid sample testing device of claim 1, wherein the medium comprises a medium which permits specific pathogens to grow on the medium. 7. The fluid sample testing device of claim 6, wherein the medium comprises a medium which permits specific uropathogens to grow on the medium.

8. The fluid sample testing device of claim 7, wherein the uropathogen specific medium is specific for primary causative agents of urinary tract infections selected from the group consisting of Escherichia coli, Kebsiella spp. , Enterobacter sp. , Serratia spp. , Proteus spp., Citrobacter spp., Acinetobacter spp., Providencia spp. , Morganella morganii, and Salmonella enterica.

9. The fluid sample testing device of claim 1, wherein the growth of microorganisms is indicated by a color change of the medium in said sample receiving members.

10. The fluid sample testing device of claim 9, wherein one or more, but not all, of the one or more sample receiving members further comprise an antimicrobial agent

11. The fluid sample testing device of claim 10, wherein the antimicrobial agent comprises at least one of amoxicillin, clavulanic acid/amoxicillin, or enrofloxacin.

12. The fluid sample testing device of claim 1 further comprising a negative control member.

13. The fluid sample testing device of claim 12, wherein the negative control member is sealed by a foil wrapper.

14. The fluid sample testing device of claim 13 further comprising:

a sterile pouch for housing the test strip with the one or more separate sample receiving members and the negative control member prior to testing.

15. The fluid sample testing device of claim 1, wherein the sample receiving members are wells each comprising an opening suitable to permit the fluid sample to contact the medium when the test strip is contacted with the fluid sample. 16. The fluid sample testing device of claim 1, wherein the sample receiving member is an absorbent pad.

17. A method for testing a fluid sample comprising:

providing a fluid sample testing kit comprising:

a test strip;

a container configured to receive and isolate the test strip; placing a fluid sample in the container;

inserting the test strip into the container, whereby the test strip is not fully immersed in the fluid sample;

causing the fluid sample to contact the one or more receiving members; and

analyzing the test strip to determine if the microorganism is present in the fluid sample.

18. The method of claim 17, wherein the fluid sample is urine.

19. The method of claim 18, wherein the medium comprises a medium which permits specific uropathogens to grow on the medium.

20. The method of claim 19, wherein the uropathogen specific medium is specific for primary causative agents of urinary tract infections selected from the group consisting of Escherichia coli, Kebsiella spp., Enterobacter sp., Serratia spp., Proteus spp., Citrobacter spp., Acinetobacter spp., Providencia spp. , Morganella morganii, and

Salmonella enterica.

21. The method of claim 17, wherein a color change in at least one sample receiving member as a result of carrying out said method indicates the microbial organism is present in the fluid sample.

22. The method of claim 17 further comprising:

providing a negative control member attached to the test strip and comprising the medium, said negative control member being sealed to prevent contact between the medium therein and the fluid sample contacting the test strip; and

determining, during said analyzing, if the microorganism is present in the fluid sample based on a comparison of the medium in the one or more sample receiving members and the medium in the negative control member.

23. The method of claim 17, wherein one or more, but not all, of the one or more sample receiving members further comprise an antimicrobial agent

24. The method of claim 23 further comprising:

determining, based on said analyzing, one or more treatment options based on a comparison of the medium in the sample receiving members without said antimicrobial agent and the medium in the sample receiving members comprising said antimicrobial agent.

25. The method of claim 23, wherein the antimicrobial agent is selected from the group consisting of amoxicillin, clavulanic acid/amoxicillin, and enrofloxacin.

26. The method of claim 24, wherein at least some of the sample receiving members containing an antimicrobial agent have different antimicrobial agents, said method further comprising:

administering the antimicrobial agent best suited for use in treatment to the subject from which the sample came.

27. The method of claim 17, wherein the container comprises an elongate tube having an opening at one end and being closed at an opposite end

28. The method of claim 27, wherein the fluid sample testing kit further comprises a cap structure configured to seal the opening of the tube, said method further comprising:

sealing the opening of the tube with said cap structure after said inserting and before said causing, wherein said causing is carried out by inverting the sealed tube one or more times.

29. The method of claim 17, wherein the sample receiving members are wells comprising an opening suitable to permit the fluid sample to contact the medium when the test strip is contacted with the fluid sample.

30. The method of claim 17, wherein the sample receiving member is an absorbent pad.

31. A fluid sample testing device comprising:

a test strip;

a negative control member attached to the test strip and comprising the medium, said negative control member being sealed to prevent contact between the medium therein and the fluid sample contacting the test strip. 32. The fluid sample testing device of claim 31, wherein the medium comprises a medium which permits specific pathogens to grow on the medium.

33. The fluid sample testing device of claim 32, wherein the medium comprises a medium which permits specific uropathogens to grow on the medium.

34. The fluid sample testing device of claim 33, wherein the uropathogen specific medium is specific for primary causative agents of urinary tract infections selected from the group consisting of Escherichia coli, Kebsiella spp., Enterobacter sp., Serratia spp., Proteus spp., Citrobacter spp., Acinetobacter spp., Providencia spp. , Morganella morganii, and Salmonella enterica.

36. The fluid sample testing device of claim 31, wherein the growth of microorganisms is indicated by a color change of the medium in said sample receiving members.

37. The fluid sample testing device of claim 36, wherein one or more, but not all, of the one or more sample receiving members further comprise an antimicrobial agent

38. The fluid sample testing device of claim 37, wherein the antimicrobial agent comprises at least one of amoxicillin, clavulanic acid/amoxicillin, or enrofloxacin.

39. The fluid sample testing device of claim 31, wherein the negative control member is sealed by a foil wrapper.

40. The fluid sample testing device of claim 31 further comprising:

41. The fluid sample testing device of claim 31, wherein the sample receiving member is a well comprising an opening suitable to permit the fluid sample to contact the medium when the test strip is contacted with the fluid sample.

42. The fluid sample testing device of claim 31, wherein the sample receiving member is an absorbent pad.

43. A method for testing a fluid sample comprising:

providing a fluid sample testing kit comprising:

a test strip;

causing the fluid sample to contact the one or more receiving members; and

44. The method of claim 43, wherein the fluid sample is urine.

45. The method of claim 44, wherein the medium comprises a medium which permits specific uropathogens to grow on the medium.

46. The method of claim 45, wherein the uropathogen specific medium is specific for primary causative agents of urinary tract infections selected from the group consisting of Escherichia coli, Kebsiella spp., Enterobacter sp., Serratia spp., Proteus spp., Citrobacter spp. , Acinetobacter spp. , Providencia spp. , Morganella morganii, and

Salmonella enterica.

47. The method of claim 43, wherein a color change in at least one sample receiving member as a result of carrying out said method indicates the microorganism is present in the fluid sample.

48. The method of claim 43 further comprising:

49. The method of claim 43, wherein one or more, but not all, of the one or more sample receiving members further comprise an antimicrobial agent 50. The method of claim 49 further comprising:

51. The method of claim 49, wherein the antimicrobial agent is selected from the group consisting of amoxicillin, clavulanic acid/amoxicillin, and enrofloxacin.

52. The method of claim 51, wherein at least some of the sample receiving members containing an antimicrobial agent have different antimicrobial agents, said method further comprising:

administering the antimicrobial agent best suited for use in treatment to the subject from which the sample came. 53. The method of claim 43, wherein the causing the fluid sample to contact the one or more receiving members comprises:

inserting the test strip into the fluid sample to provide contact between the receiving members and the fluid sample; and removing the test strip from the fluid sample.

54. The method of claim 43, wherein the causing the fluid sample to contact the one or more receiving members comprises:

exposing the test strip to a stream of the fluid sample to provide contact between the receiving members and the fluid sample.

55. The method of claim 43 further comprising:

storing the test strip in a container after causing the fluid sample to contact the one or more receiving members, wherein the test strip is no longer in contact with the fluid sample.

56. The method of claim 43, wherein the sample receiving member is a well comprising an opening suitable to permit the fluid sample to contact the medium when the test strip is contacted with the fluid sample.

57. The method of claim 43, wherein the sample receiving member absorbent pad.