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Journal of Clinical Microbiology, May 2000, p. 1953-1958, Vol. 38, No. 5
0095-1137/00/$04.00+0

Rapid Identification of Yersinia enterocolitica in Blood by the 5' Nuclease PCR Assay

Keya Sen*

Division of Emerging and Transfusion Transmitted Diseases, Center for Biologics Evaluation and Research, Food and Drug Administration, Rockville, Maryland 20852

Received 14 December 1999/Returned for modification 20 January 2000/Accepted 23 February 2000

Yersinia enterocolitica accounts for 50% of the clinical sepsis episodes caused by the transfusion of contaminated red blood cells. A 5' nuclease TaqMan PCR assay was developed to detect Y. enterocolitica in blood. Primers and a probe based on the nucleotide sequence of the 16S rRNA gene from Y. enterocolitica were designed. Whole-blood samples were spiked with various numbers of Y. enterocolitica cells, and total chromosomal DNA was extracted. When the TaqMan PCR assay was performed, as few as six bacteria spiked in 200 µl of blood could be detected. The assay was specific and did not detect other Yersinia species. The TaqMan assay is easy to perform, takes 2 h, and has the potential for use in the rapid detection of Y. enterocolitica contamination in stored blood units.

* Mailing address: Division of Emerging and Transfusion Transmitted Diseases, Center for Biologics Evaluation and Research, Food and Drug Administration, HFM-320, 1401 Rockville Pike, Rockville, MD 20852. Phone: (301) 594-6752. Fax: (301) 594-6989. E-mail: Senk{at}cber.fda.gov.

Journal of Clinical Microbiology, May 2000, p. 1953-1958, Vol. 38, No. 5
0095-1137/00/$04.00+0


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