Growth-independent detection of cells
Disclosed herein are various methods, systems, and compositions for the growth independent detection of cells such as microorganisms including bacteria. While existing cellular detection methodologies benefit from cell growth, the methods, systems, and compositions disclosed herein demonstrate embodiments that are independent of cell growth while still allowing for cell-based detection.
1. A sample or cell culture comprising a plurality of non-replicative transduction particles (NRTPs) and one or more microorganisms of interest, wherein the plurality of NRTPs are produced using a silent mutation/complementation method or a deletion/complementation method, wherein said methods do not produce bacteriophages or NRTPs that contain a viral genome, and wherein the plurality of NRTPs comprises a reporter nucleic acid sequence, and wherein the growth rate of the one or more microorganisms of interest is less than logarithmic phase.
2. The sample or cell culture of claim 1 , further comprising an antimicrobial agent.
3. The sample or cell culture of claim 1 , wherein the one or more microorganisms of interest is in stationary phase; or wherein the one or more microorganisms of interest is undergoing no growth.
4. The sample or cell culture of claim 1 , wherein the growth rate of the one or more microorganisms of interest is negative or homeostatic.
5. The sample or cell culture of claim 1 , wherein the one or more microorganisms of interest comprises a Methicillin Resistant Staphylococcus aureus (MRSA) cell, Staphylococcus aureus, Staphylococcus spp., Enterobacteriaceae, Enterococcus spp. Streptococcus spp., Acinetobacter spp., or Pseudomonas spp.
6. The sample or cell culture of claim 1 , wherein the sample is a clinical sample.
7. The sample or cell culture of claim 1 , further comprising a fatty aldehyde bacterial luciferase substrate reagent, optionally wherein the reagent is tridecanal.
8. The sample or cell culture of claim 2 , wherein the antimicrobial agent is cefoxitin, a β-lactam, an extended-spectrum β-lactam, an Aminoglycoside, an Ansamycin, a Carbacephem, Carbapenems, any generation of Cephalosporin, a Glycopeptide, a Lincosamide, a Lipopeptide, a Macrolide, a Monobactam, a Nitrofuran, an Oxazolidonone, a Penicillin, a Polypeptide, a Quinolone, a Fluoroquinolone, a Streptogramin, a Sulfonamide, a Tetracycline, a Rifampicin, a mycobacterial antibiotic, Chloramphenicol, and/or Mupirocin.
9. The sample or cell culture of claim 1 , wherein the reporter nucleic acid sequence encodes a detectable or selectable marker.