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Journal of Clinical Microbiology, October 2003, p. 4873-4875, Vol. 41, No. 10
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.10.4873-4875.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Detection of Yersinia pestis in Sputum by Real-Time PCR

Caroline Loïez,1 Stéphanie Herwegh,1 Frédéric Wallet,1,2 Sylvie Armand,1 Françoise Guinet,3 and René J. Courcol1,2*

Laboratoire de Bactériologie-Hygiène, Centre Hospitalier Régional Universitaire de Lille,1 Equipe Mixte Inserm—Université 0364, Institut de Biologie de Lille, Lille,2 Centre National de Référence des Yersinia, Institut Pasteur, Paris, France3

Received 6 February 2003/ Returned for modification 19 March 2003/ Accepted 10 July 2003

A 5' nuclease PCR assay for detection of the Yersinia pestis plasminogen activator (pla) gene in human respiratory specimens with simulated Y. pestis infection was developed. An internal positive control was added to the reaction mixture in order to detect the presence of PCR inhibitors that are often found in biological samples. The assay was 100% specific for Y. pestis. In the absence of inhibitors, a sensitivity of 102 CFU/ml of respiratory fluid was obtained. When inhibitors were present, detection of Y. pestis DNA required a longer sample treatment time and an initial concentration of bacteria of at least 104 CFU/ml. The test's total turnaround time was less than 5 h. The assay described here is well suited to the rapid diagnosis of pneumonic plague, the form of plague most likely to result from a bioterrorist attack.

* Corresponding author. Mailing address: Laboratoire de Bactériologie-Hygiène, Hôpital A. Calmette, F-59037, Lille cedex, France. Phone: (33) 320 444 908. Fax: (33) 320 444 895. E-mail: rcourcol{at}chru-lille.fr.

Journal of Clinical Microbiology, October 2003, p. 4873-4875, Vol. 41, No. 10
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.10.4873-4875.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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