Modification of sample preparation to differentiate live and dead bacteria by polymerase chain reaction assay

摘要

The invention relates to a method of determining whether a live microbe, such as bacteria, is present in a test sample.

主权项

1. A method of detecting the presence of a live microbe in a test sample suspected of containing the live microbe, the method comprising:
a) isolating the microbe from the known culture; b) adding 5, 5′-(6, 22-dioxo-11, 14, 17-trioxa-7, 21-diazaheptacosane-1, 27-diyl) bis (3, 8-diamino-6-phenylphenanthridin-5-ium) iodide dye to the isolated microbe from step (a); c) extracting DNA from the microbe after step (b); d) performing PCR on the DNA from step (c); e) analyzing PCR results from step (d) for the presence or absence of amplified DNA using real time PCR and further gel electrophoresis confirmation; f) correlating the presence of amplified DNA from step (e) with the presence of live bacteria in the test sample.