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

Q. Lou, S. K.F. Chong, J. F. Fitzgerald, J. A. Siders, S. D. Allen, C. H. Lee

Research output: Contribution to journalArticle

31 Scopus citations

Abstract

We have developed a novel method for the preparation of fecal specimens for PCR assays. Approximately 100 mg of solid stool or 200 μl 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.

Original languageEnglish (US)
Pages (from-to)281-283
Number of pages3
JournalJournal of clinical microbiology
Volume35
Issue number1
StatePublished - Jan 1 1997

ASJC Scopus subject areas

  • Microbiology (medical)

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    Lou, Q., Chong, S. K. F., Fitzgerald, J. F., Siders, J. A., Allen, S. D., & Lee, C. H. (1997). Rapid and effective method for preparation of fecal specimens for PCR assays. Journal of clinical microbiology, 35(1), 281-283.