Reagent cartridge and methods for detection of cells
An apparatus includes a housing and an actuator. The housing, which defines a reagent volume that can receive a reagent container, can be removably coupled to a reaction chamber. The housing includes a puncturer that defines a transfer pathway in fluid communication with the reagent volume. A delivery portion of the housing defines a delivery pathway between the transfer pathway and the reaction chamber when the housing is coupled to the reaction chamber. The actuator has a plunger portion disposed within the reagent volume. An engagement portion of the actuator can be manipulated to move the plunger portion within the reagent volume to deform the reagent container. The puncturer can pierce a frangible portion of the reagent container to convey a reagent from the reagent container into the reaction chamber via the transfer pathway and/or the delivery pathway.
1. A method of determining the presence of an antibiotic resistant cell in a biological sample, comprising:
disposing the biological sample into a reaction chamber, the reaction chamber packaged to contain an antibiotic formulated to mix with the biological sample to form an assay media;
mixing with the assay media in the reaction chamber a plurality of transduction particles associated with a cell phenotype, the plurality of transduction particles comprising a nucleic acid molecule formulated to cause the cell phenotype to produce a plurality of reporter molecules capable of generating a detectable signal, the antibiotic formulated to suppress the detectable signal in the cell phenotype;
receiving, by a first detector of a detection instrument, a first signal indicating the presence of the antibiotic;
maintaining, for a period of about eight hours or less, the assay media and the plurality of transduction particles within the detection instrument to express the plurality of reporter molecules when the cell phenotype is present in the biological sample; and
receiving, by a second detector of the detection instrument, a second signal indicating a quantity of the plurality of reporter molecules thereby determining the presence of the antibiotic resistant cell.
2. The method of claim 1 , wherein the antibiotic is a dried antibiotic.
3. The method of claim 1 ; wherein the antibiotic is adhered to an inner surface of the reaction chamber.
4. The method of claim 1 , wherein the cell phenotype comprises a bacteria phenotype that is resistant to the antibiotic individually or in combination with another antibiotic.
5. The method of claim 4 ; wherein the antibiotic is any one of Beta-lactams, extended-spectrum beta-lactams, Aminoglycosides, Ansamycins, Carbacephem, Carbapenems, any generation of Cephalosporins, Glycopeptides, Lincosamides, Lipopeptide, Macrolides, Monobactams, Nitrofurans, Oxazolidonones, Penicillins, Polypeptides, Quinolones, Fluoroquinolones, Sulfonamides, Tetracyclines, mycobacterial antibiotics, Chloramphenicol, Mupirocin.
6. The method of claim 1 , wherein the cell phenotype comprises a bacteria phenotype that is resistant to at least one of cefoxitin, vancomycin, teicoplainin, ampicillin/sulbactam, ciprofloxacin, meropenem, ceftazidime, ceftriaxone, piperacillin/tazobactam, or gentamicin.
7. The method of claim 1 , wherein:
the antibiotic comprises a colorant, the antibiotic formulated to suppress production of the plurality of reporter molecules in the cell phenotype; and
the first signal is associated with the colorant and indicates the presence of the antibiotic.
8. The method of claim 1 , wherein the first signal is associated with a volume of the assay media within the reaction chamber.
9. The method of claim 1 ; wherein a magnitude of the second signal is independent from the quantity of the plurality of transduction particles when the quantity is above a predetermined quantity.
10. The method of claim 1 , wherein the plurality of transduction particles is engineered to be incapable of either of lytic or lysogenic replication.
11. The method of claim 1 , wherein the plurality of transduction particles is derived from a bacteriophage.
12. The method of claim 1 , further comprising:
disposing, after the maintaining, a substance into the assay media, the substance formulated to react with the plurality of reporter molecules to generate the second signal.
13. A method of determining the presence of an antibiotic resistant target cell in a biological sample, comprising:
receiving a container containing a swab and a transport media, the swab comprising a shaft having a collection portion constructed from anon-wound material, the transport media comprising the biological sample released from the collection portion;
transferring the transport media and the biological sample into a reaction chamber;
mixing with the transport media and the biological sample within the reaction chamber a plurality of transduction particles associated with the antibiotic resistant target cell, the plurality of transduction particles comprising a nucleic acid molecule formulated to cause the antibiotic resistant target cell to produce a plurality of reporter molecules capable of generating a detectable signal;
maintaining the mixture of the transport media, the biological sample, and the plurality of transduction particles at a temperature of at least 20 C for a period of about eight hours or less to express the plurality of reporter molecules when the antibiotic resistant target cell is present in the biological sample; and
receiving, by a detector of a detection instrument, the detectable signal indicating a quantity of the plurality of reporter molecules, thereby determining the presence of the antibiotic resistant cell.
14. The method of claim 13 , wherein the plurality of transduction particles is non-replicative.
15. The method of claim 13 , wherein the plurality of transduction particles is devoid of a reporter molecule of the plurality of reporter molecules.
16. The method of claim 13 , wherein a reporter molecule from the plurality of reporter molecules is any one of a bacterial luciferase, an eukaryotic luciferase, a fluorescent protein, an enzyme suitable for colorimetric detection, a protein suitable for immunodetection, a peptide suitable for immunodetection or a nucleic acid that function as an aptamer or that exhibits enzymatic activity.
17. The method of claim 13 , wherein the maintaining is performed for a period of about two hours or less.
18. The method of claim 13 , wherein the collection portion is constructed from a foam material.
19. The method of claim 13 , wherein:
the reaction chamber contains a lyophilized reagent tablet formulated to mix with the transport media, the lyophilized reagent tablet comprising an antibiotic.
20. A method of determining the presence of an antibiotic resistant target cell in a biological sample, comprising:
receiving a container containing a swab and a transport media, the swab comprising a shaft having a collection portion constructed from non-wound material, the transport media comprising the biological sample released from the collection portion;
transferring the transport media and the biological sample into a reaction chamber; and
actuating a detection instrument to cause the detection instrument to:
mix with the transport media and the biological sample within the reaction chamber a plurality of transduction particles associated with the antibiotic resistant target cell; the plurality of transduction particles comprising a nucleic acid molecule formulated to cause the antibiotic resistant target cell to produce a plurality of reporter molecules capable of generating a detectable light signal;
maintain within the detection instrument the mixture of the transport media and the plurality of transduction particles at a temperature of at least 20 C for a period of about eight hours or less to express the plurality of reporter molecules when the antibiotic resistant target cell is present in the biological sample; and
receive, by a detector of the detection instrument, the detectable light signal indicating a quantity of the plurality of reporter molecules thereby determining the presence of the antibiotic resistant cell.