Rapid and effective method for preparation of fecal specimens for PCR assays.

Journal of Clinical Microbiology
Q LouC H Lee

Abstract

We have developed a novel method for the preparation of fecal specimens for PCR assays. Approximately 100 mg of solid stool or 200 microliters of liquid fecal sample was thoroughly suspended in 1 ml of water. Fecal debris was removed by low-speed centrifugation (2,800 x g for 2 min). The supernatant was then boiled for 10 min in a water bath and further clarified by high-speed centrifugation (12,000 x g for 5 min). Fifty microliters of the clarified supernatant was then purified by Sepharose CL-6B spin column chromatography, and a portion of the purified supernatant was used for PCR. By this method, stools containing enterotoxigenic Escherichia coli H10407 were amplified by colonization factor antigen I fimbrial gene PCR, with a sensitivity of 100 organisms per reaction. The method was also effective for processing stool specimens for Clostridium difficile toxin A and B gene PCRs. This method is rapid, effective, and simple to perform and will improve the applications of PCR to stool specimens for diagnostic purposes.

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Citations

Apr 21, 2001·FEMS Microbiology Letters·A S MikoszaD J Hampson
Apr 9, 2008·Avian Pathology : Journal of the W.V.P.A·L BanoS F Bilgili
Nov 26, 2009·International Journal of Microbiology·Magdalena JacobsonAnna Aspan
Oct 16, 2007·Applied and Environmental Microbiology·M RibièreJ-P Faucon
Dec 3, 1998·Applied and Environmental Microbiology·A Tilsala-Timisjärvi, T Alatossava
Feb 11, 1998·Clinical Microbiology Reviews·J P Nataro, J B Kaper
May 13, 2011·Journal of the American Chemical Society·B Scott FergusonH Tom Soh

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