Rapid detection of replicating cells
OwnerDon Strausinventor
Life of this patent
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Abstract
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.
Claims99
A method for detecting microcolonies of living target cells in a sample, said method comprising the steps of:
- (a) depositing 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;
- and
- (c) detecting said one or more microcolonies;
- wherein the longest linear dimension of said detection area is greater than 1 mm;
- said one or more microcolonies have a measurement of less than 50 microns in at least two orthogonal dimensions;
- said detecting does not entail magnification of more than 5×;
- said detecting detects a property of said one or more microcolonies that does not depend on the addition of a signalling moiety or category binding molecule;
- and said cells in said one or more microcolonies remain competent to replicate following said detecting.
Dependencies are inferred from the claim text.