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US 9,434,976 B2Granted
Modification of sample preparation to differentiate live and dead bacteria by polymerase chain reaction assay
AssigneesTuskegee University, Tsegaye Habtermariam
InventorsTeshome Yehualaeshet, Temesgen Samuel, Woubit Abdela, Tsegaye Habtemariam
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Abstract
The invention relates to a method of determining whether a live microbe, such as bacteria, is present in a test sample.
Claims9
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.
3 more elements
Dependencies are inferred from the claim text.