Rapid detection of replicating cells

摘要

The invention enables efficient, rapid, and sensitive enumeration of living cells by detecting microscopic colonies derived from in situ cell division using large area imaging. Microbial enumeration tests based on the invention address an important problem in clinical and industrial microbiology—the long time needed for detection in traditional tests—while retaining key advantages of the traditional methods based on microbial culture. Embodiments of the invention include non-destructive aseptic methods for detecting cellular microcolonies without labeling reagents. These methods allow for the generation of pure cultures which can be used for microbial identification and determination of antimicrobial resistance.

主权项

1. A method for detecting living target cells in a sample, said method comprising the steps of:
(a) providing living target cells present in said sample in a detection zone comprising a detection area at a density of less than 100 target cells per mm2 of the detection area, wherein within said detection zone said cells are randomly dispersed and immobilized; (b) allowing the formation of one or more microcolonies of said target cells by in situ replication, (c) labeling said one or more microcolonies with a labeling reagent; and (d) detecting said one or more microcolonies by detecting the signal generated by said labeling reagent;wherein the longest linear dimension of said detection area is greater than 1 mm; said one or more microcolonies have a mean measurement of less than 50 microns in at least two orthogonal dimensions; wherein said detecting does not entail magnification of more than 5×; and wherein said cells in said one or more microcolonies remain competent to replicate following said detecting.