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Journal of Clinical Microbiology, August 1998, p. 2284-2288, Vol. 36, No. 8
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

5' Nuclease PCR Assay To Detect Yersinia pestis

James A. Higgins,1 John Ezzell,1 B. Joseph Hinnebusch,2 Michelle Shipley,1 Erik A. Henchal,1 and M. Sofi Ibrahim1,*

Diagnostic Systems Division, United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, Maryland 21702-5011,1 and Laboratory of Microbial Structure and Function, Rocky Mountain Laboratories, National Institute of Allergy and Infectious Diseases, Hamilton, Montana 598402

Received 17 November 1997/Returned for modification 13 February 1998/Accepted 10 April 1998

The 5' nuclease PCR assay uses a fluorescently labeled oligonucleotide probe (TaqMan) to rapidly detect and quantitate DNA templates in clinical samples. We developed a 5' nuclease PCR assay targeting the plasminogen activator gene (pla) of Yersinia pestis. The assay is species specific, with a detection threshold of 2.1 × 105 copies of the pla target or 1.6 pg of total cell DNA. The assay detected Y. pestis in experimentally infected Xenopsylla cheopis fleas and in experimentally infected monkey blood and oropharyngeal swabs. The TaqMan assay is simple to perform and rapid and shows promise as a future field-adaptable technique.

* Corresponding author. Mailing address: Diagnostic Systems Division, USAMRIID, 1425 Porter St., Ft. Detrick, MD 21702-5011. Phone: (301) 619-4721. Fax: (301) 619-2492. E-mail: sibrahim{at}detrick.army.mil.

Journal of Clinical Microbiology, August 1998, p. 2284-2288, Vol. 36, No. 8
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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