Device and method for identifying microbes and counting microbes and determining antimicrobial sensitivity

Abstract

A method of determining antimicrobial activity of an agent can include providing a well, wherein the well contains at least one antimicrobial agent, the well further including at least two electrodes. A sample of a microbe can be added into the well and a voltage pulsed between the electrodes. An electrical property can be sampled and recorded. In another aspect, a method of identifying at least one microbe includes taking a sample containing the at least one microbe, isolating the at least one microbe from the sample, dividing the at least one microbe into at least one well, wherein each well contains at least one antimicrobial agent and at least two electrodes. A voltage is pulsed between the at least two electrodes, an electrical property is sampled during the pulsing and recorded. In another aspect, a diagnostic device for detecting at least one microbe is presented.

What is claimed is: 
     
         1 . A method of determining a count of microbes in a sample, the method comprising:
 filtering the sample to separate the at least one microbe from the sample,   immersing the at least one microbe in an analyte to form an immersion,   incubating the immersion for a specific time,   dividing the immersion into at least one wells,   measuring an electrical property in the wells for a sample-duration, and   correlating the electrical property to a count as a function of time.   
     
     
         2 . The method according to  claim 1 , further comprising adding at least one bacteriophage to at least one of the wells before measuring the electrical property. 
     
     
         3 . The method according to  claim 1 , wherein the sample-duration is at is about 1 second, about 5 seconds, about 10 seconds, about 20 seconds, about 30 seconds, about 40 seconds, about 50 seconds, about 60 seconds. 
     
     
         4 . The method according to  claim 1 , wherein the specific time for the incubating is between 1 millisecond and 5 minutes, between 0.5 second and 2 minutes, between 1 second and 1 minute, between 1 hour and 8 hours. 
     
     
         5 . The method according to  claim 1 , wherein the method determines a first count for a first microbe and a second count for a second microbe. 
     
     
         6 . A method of determining antimicrobial resistance of a microbe, comprising:
 adding a sample of at least one microbe into a well containing at least one antimicrobial, and adding a sample of at least one microbe into a control well   measuring an electrical property in the well for a sample-duration as a function of time.   
     
     
         7 . The method according to  claim 6  where the sample-duration is at least one hour and not more than six hours. 
     
     
         8 . The method according to  claim 6 , wherein the at least one microbe is selected from  Aerobacter, Bacillus, Bordetella, Brucella, Campylobacter, Chlamydia, Chromobacterium, Clostridium, Corynebacterium, Enterobacter, Escherichia, Haemophilus, Klebsiella, Listeria, Mycobacterium, Mycoplasma, Neisseria, Pneumococcus, Proteus, Pseudomonas, Providencia, Salmonella, Serratia, Shigella, Staphylococcus, Streptococcus, Vibrio, Yersinia, Acinetobacter, Bacteroides, Bifidubacterium, E. kenella corrodens, Gardnerella vaginalis, Mobiluncus, Proteobacteria, Desulfobacterales, Desulfovibrionales, Syntrophobacterales, Thermodesulfobacteria, Nitrospirae,  gram positive Peptococcaceae, Archaea,  Archaeoglobus , or any combinations thereof. 
     
     
         9 . The method according to  claim 6 , wherein the at least one agent is selected from  Actinomyces  phages,  Bacillus  phage Φ29, bacteriophage M102, bacteriophage e10, bacteriophage f1, bacteriophage λ, bacteriophage PI, spherical phage PhiX174, spherical phage G4, spherical phage S13, bacteriophage T1, bacteriophage T2, bacteriophage T3, bacteriophage T4, bacteriophage T5, bacteriophage T6, bacteriophage T7, ssRNA bacteriophages MS2, ssRNA bacteriophages R17, ssRNA bacteriophages f2, ssRNA bacteriophages Q beta,  S. mutans  phages, and any combinations thereof. 
     
     
         10 . The method according to any one of the  claim 6 , wherein the well has a holding capacity between about 10 μL to about 2 mL. 
     
     
         11 . The method according to  claim 6 , wherein the electrical property is selected from conductance, resistance, voltage, amperage, capacitance, impedance, inductance, and any combinations thereof. 
     
     
         12 . The method according to  claim 6 , wherein the sample is taken from urine, blood, sweat, mucus, saliva, semen, vaginal secretion, vomit, tears, sebum, pleural fluid, peritoneal fluid, gastric juice, earwax, cerebrospinal fluid, breast milk, endolymph, perilymph, aqueous humor, vitreous humor, biomass and any combinations thereof. 
     
     
         13 . A diagnostic device of detecting at least one microbe, comprising:
 a first unit and a second unit; the first unit is stackable into the second unit;
 wherein the first unit is a diagnostic unit comprising at least one well, the at least one well having electrodes contacting the inside and the outside of the at least one well; 
 wherein the second unit is a reader unit comprising a connector section for the electrodes of the diagnostic unit; a fluidic system consisting of one-way valves and a port for pressurizing the fluidic system; 
 wherein the second unit is a reader unit comprising a connector section for the pressurizing. 
   
     
     
         14 . The diagnostic device according to  claim 13 , wherein the first unit further comprising a sample holder and filter unit, the sample holder and filter unit being in fluidic communication. 
     
     
         15 . The diagnostic devices according to  claim 13 , wherein the electrodes comprise a non-oxidizing material. 
     
     
         16 . The diagnostic devices according to  claim 15 , wherein the non-oxidizing material is selected from metals, nonmetals, polymers, composites, resists, resins, carbon nano-tubes, plastics, or any combinations thereof. 
     
     
         17 . The diagnostic devices according to  claim 13 , wherein the electrodes comprises copper covered with graphene. 
     
     
         18 . The diagnostic devices according to  claim 16 , wherein the electrodes comprise copper, gold, nickel, or any combination thereof. 
     
     
         19 . The diagnostic device according to  claim 13 , wherein the at least one antimicrobial is selected from aminoglycosides, amphenicols, ansamycins, beta-lactams, lincosamides, macrolides, polypeptide antibiotics, tetracyclines, cycloserine, mupirocin, tuberin, 2,4-diaminopyrimidines, nitrofurans, quinolones, sulfonamides, sulfones, clofoctol, hexedine, methenamine, nitroxoline, taurolidine, and xibernol. 
     
     
         20 . The diagnostic device according to  claim 13 , wherein the at least one antimicrobial is selected from amikacin, azlocillin, carbencillin, cefaclor, cefemandole, cefonicid, cefotaxime, cefoperazone, cefoxitin, ceftizoxime, ceftriaxzone, ciprofloxacin, clindamycin, gatifloxacin, gemifloxacin, gentamicin, kanamycin, linezolid, mecillinam, meropenem, methicillin, metronidazole, mezlocillin, minocyclin, moxifloxacin, nafcillin, netilmycin, oxacillin, penicillin, piperacillin, quinupristin-dalfopristin, sparfloxacin, sulbactam, tazobactam, teicoplanin, tetracyclines, tobramycin, trimethoprim, trospectomycin and vancomycin.