Antimicrobial cartridges and processes for multiplexed antimicrobial susceptibility testing
The present invention provides, among other things, cartridges for multiplexed antimicrobial susceptibility testing (AST), single cartridges useful for both AST and quality control of AST, and systems and methods relating thereto.
1. A cartridge for performing antimicrobial susceptibility testing of microorganisms, comprising:
a plurality of reservoirs arranged in a series of rows and columns, each reservoir containing an antimicrobial agent; and
a plurality of reservoir sets, each reservoir set comprising a series of adjacent reservoirs along a row of the cartridge and comprising a dilution series of the same antimicrobial agent;
wherein the columns of reservoirs within the cartridge are divided into at least two regions, wherein each region comprises a group of columns for receiving a quality control organism that is different from a quality control organism received by a different region, and each region has at least two reservoir sets for assessing the antimicrobial susceptibility of the quality control organism received in that region;
wherein the reservoir sets within each region comprise different antimicrobial agents from each other; and
wherein the cassette is adapted to receive a patient sample or first and second quality control organisms, the first quality control organism being applied to a reservoir set within a first region of the at least two regions, and the second quality control organism being applied to a reservoir within a second region of the at least two regions.
2. The cartridge of claim 1 , wherein each column region comprises at least four reservoir sets.
3. The cartridge of claim 1 , wherein each column region comprises at least eight reservoir sets.
4. The cartridge of claim 1 , comprising at least 384 reservoirs.
5. The cartridge of claim 1 , wherein each dilution series comprises sufficient dilution extents to include a minimum inhibitory concentration of a quality control organism applied to that dilution series.
6. The cartridge of claim 1 , wherein two or more reservoirs of the cartridge comprise a sufficient quantity of an antimicrobial agent to inhibit the growth of a plurality of gram-negative microorganisms.
7. The cartridge of claim 1 , in which one or more of the antimicrobial agents is selected from the group consisting of Amikacin, Amikacin-fosfomycin, Amoxicillin, Amoxicillin-clavulanate, Ampicillin, Ampicillin-sulbactam, Azithromycin, Azithromycin-Avibactam, Azlocillin, Aztreonam, Aztreonam-avibactam, Besifloxacin, Biapenem, Cadazolid, Carbenicillin, Cefaclor, Cefamandole, Cefazolin, Cefdinir, Cefditoren, Cefepime, Cefepime-tazobactam, Cefetamet, Cefiderocol, Cefixime, Cefmetazole, Cefonicid, Cefoperazone, Cefotaxime, Cefotetan, Cefoxitin, Ceftolozane-tazobactam, Cefpodoxime, Cefprozil, Ceftaroline, Ceftaroline-avibactam, Ceftazidime, Ceftazidime-avibactam, Ceftibuten, Ceftizoxime, Ceftobiprole, Ceftolozane-tazobactam, Ceftriaxone, Cefuroxime, Cephalothin, Chloramphenicol, Cinoxacin, Ciprofloxacin, Clarithromycin, Clinafloxacin, Clindamycin, Colistin, Dalbavancin, Daptomycin, Delafloxacin, Dirithromycin, Doripenem, Doxycycline, Enoxacin, Eravacycline, Ertapenem, Erythromycin, Faropenem, Fidaxomicin, Finafloxacin, Fleroxacin, Fosfomycin, Fusidic acid, Garenoxacin, Gatifloxacin, Gemifloxacin, Gentamicin, Gepotidacin, Grepafloxacin, Iclaprim, Imipenem, Imipenem-Relebactam, Kanamycin, Lefamulin, Levofloxacin, Levonadifloxacin, Linezolid, Linopristin-flopristin, Lomefloxacin, Loracarbef, Mecillinam, Meropenem, Methicillin, Mezlocillin, Minocycline, Moxalactam, Moxifloxacin, Nafcillin, Nalidixic acid, Netilmicin, Nitrofurantoin, Norfloxacin, Ofloxacin, Omadacycline, Oritavancin, Oxacillin, Penicillin, Piperacillin, Piperacillin-tazobactam, Plazomicin, Polymyxin B, Quinupristin-dalfopristin, Razupenem, Rifampin, Solithromycin, Sparfloxacin, Sulfisoxazole, Sulopenem, Tedizolid, Teicoplanin, Televancin, Telithromycin, Tetracycline, Ticarcillin, Ticarcillin-clavulanate, Tigecycline, Tobramycin, Trimethoprim, Trimethoprim-sulfamethoxazole, Trospectomycin, Vancomycin, Aculeacin A, Amphotericin B, Caspofungin, Clotrimazole, Fluconazole, Flucytosine, 5-Fluorocytosine, Griseofulvin, Itraconazole, Ketoconazole, Nystatin, Sordarin, Terbinafine, Vaborbactam-meropenem, Voriconazole and a salt or hydrate form thereof.
8. The cartridge of claim 1 , wherein one or more quality control organisms is selected from the group consisting of Staphylococcus aureus ssp. aureus (ATCC 29213); Enterococcus faecalis (ATCC 29212);
Escherichia coli (ATCC 25922);
Klebsiella pneumoniae ssp. pneumoniae (ATCC 700603);
Klebsiella pneumoniae (ATCC BAA2814);
Pseudomonas aeruginosa (ATCC 27853); E. faecalis (ATCC 51299); S. aureus (BAA-1708); S. aureus (BAA-977); S. aureus (ATCC 43300); Streptococcus pneumoniae (ATCC 49619) and Trichothecium plasmoparae (ATCC 13353).
9. The cartridge of claim 1 , wherein each dilution series is replicated two or more times on the cartridge, and the cartridge is inoculated with two or more patient samples.
10. The cartridge of claim 1 , wherein the cartridge is inoculated with a patient sample comprising a gram-negative microorganism, and at least two reservoirs in the cartridge are inoculated at a higher concentration than the other reservoirs in the cartridge.
11. A kit for performing antimicrobial susceptibility testing (AST), comprising:
a cartridge according to claim 1 ; and
instructions for performing:
(a) an AST method comprising the steps of:
inoculating the cartridge with a patient sample; and
assessing, based on a comparison of cell growth in differing antimicrobial concentrations, one of a minimum inhibitory concentration of an antimicrobial agent, and a susceptibility to an antimicrobial agent; and
(b) a quality control method comprising the steps of:
inoculating the cartridge with at least one quality control organism specified for an antimicrobial agent present on the cartridge; and
assessing, based on a comparison of cell growth in differing antimicrobial concentrations and a normal range for the quality control organism, whether a minimum inhibitory concentration of an antimicrobial agent is within the normal range for the quality control organism.
12. The kit of claim 11 , wherein the quality control method comprises the step of simultaneously applying the first and second quality control organism to the cartridge.
13. The kit of claim 11 , wherein each column region comprises at least four reservoir sets.
14. The kit of claim 11 , wherein each column region comprises at least eight reservoir sets.
15. The kit of claim 11 , wherein the cassette comprises at least 384 reservoirs.
16. The kit of claim 11 , wherein each dilution series comprises sufficient dilution extents to include a minimum inhibitory concentration of a quality control organism applied to that dilution series.
17. The kit of claim 11 , wherein two or more reservoirs of the cartridge comprise a sufficient quantity of an antimicrobial agent to inhibit the growth of a plurality of gram-negative microorganisms.
18. The kit of claim 11 , in which one or more of the antimicrobial agents are selected from the group consisting of Amikacin, Amikacin-fosfomycin, Amoxicillin, Amoxicillin-clavulanate, Ampicillin, Ampicillin-sulbactam, Azithromycin, Azithromycin-Avibactam, Azlocillin, Aztreonam, Aztreonam-avibactam, Besifloxacin, Biapenem, Cadazolid, Carbenicillin, Cefaclor, Cefamandole, Cefazolin, Cefdinir, Cefditoren, Cefepime, Cefepime-tazobactam, Cefetamet, Cefiderocol, Cefixime, Cefmetazole, Cefonicid, Cefoperazone, Cefotaxime, Cefotetan, Cefoxitin, Ceftolozane-tazobactam, Cefpodoxime, Cefprozil, Ceftaroline, Ceftaroline-avibactam, Ceftazidime, Ceftazidime-avibactam, Ceftibuten, Ceftizoxime, Ceftobiprole, Ceftolozane-tazobactam, Ceftriaxone, Cefuroxime, Cephalothin, Chloramphenicol, Cinoxacin, Ciprofloxacin, Clarithromycin, Clinafloxacin, Clindamycin, Colistin, Dalbavancin, Daptomycin, Delafloxacin, Dirithromycin, Doripenem, Doxycycline, Enoxacin, Eravacycline, Ertapenem, Erythromycin, Faropenem, Fidaxomicin, Finafloxacin, Fleroxacin, Fosfomycin, Fusidic acid, Garenoxacin, Gatifloxacin, Gemifloxacin, Gentamicin, Gepotidacin, Grepafloxacin, Iclaprim, Imipenem, Imipenem-Relebactam, Kanamycin, Lefamulin, Levofloxacin, Levonadifloxacin, Linezolid, Linopristin-flopristin, Lomefloxacin, Loracarbef, Mecillinam, Meropenem, Methicillin, Mezlocillin, Minocycline, Moxalactam, Moxifloxacin, Nafcillin, Nalidixic acid, Netilmicin, Nitrofurantoin, Norfloxacin, Ofloxacin, Omadacycline, Oritavancin, Oxacillin, Penicillin, Piperacillin, Piperacillin-tazobactam, Plazomicin, Polymyxin B, Quinupristin-dalfopristin, Razupenem, Rifampin, Solithromycin, Sparfloxacin, Sulfisoxazole, Sulopenem, Tedizolid, Teicoplanin, Televancin, Telithromycin, Tetracycline, Ticarcillin, Ticarcillin-clavulanate, Tigecycline, Tobramycin, Trimethoprim, Trimethoprim-sulfamethoxazole, Trospectomycin, Vancomycin, Aculeacin A, Amphotericin B, Caspofungin, Clotrimazole, Fluconazole, Flucytosine, 5-Fluorocytosine, Griseofulvin, Itraconazole, Ketoconazole, Nystatin, Sordarin, Terbinafine, Vaborbactam-meropenem, Voriconazole and a salt or hydrate form thereof.
19. The kit of claim 11 , wherein one or more quality control organisms are selected from the group consisting of Staphylococcus aureus ssp. aureus (ATCC 29213); Enterococcus faecalis (ATCC 29212);
Escherichia coli (ATCC 25922);
Klebsiella pneumoniae ssp. pneumoniae (ATCC 700603);
Klebsiella pneumoniae (ATCC BAA2814);
Pseudomonas aeruginosa (ATCC 27853); E. faecalis (ATCC 51299); S. aureus (BAA-1708); S. aureus (BAA-977); S. aureus (ATCC 43300); Streptococcus pneumoniae (ATCC 49619) and Trichothecium plasmoparae (ATCC 13353).
20. The kit of claim 11 , wherein each dilution series is replicated two or more times on the cartridge, and the cartridge is inoculated with two or more patient samples.
21. The kit of claim 11 , wherein the cartridge is inoculated with a patient sample comprising a gram-negative microorganism, and at least two reservoirs in the cartridge are inoculated at a higher concentration than the other reservoirs in the cartridge.