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Journal of Clinical Microbiology, September 2001, p. 3047-3051, Vol. 39, No. 9
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.9.3047-3051.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Rapid and Accurate Identification of Coagulase-Negative Staphylococci by Real-Time PCR

K. J. Edwards,1 M. E. Kaufmann,2 and N. A. Saunders1,*

Molecular Biology Unit, Virus Reference Division,1 and Laboratory of Hospital Infection,2 Central Public Health Laboratory, Colindale, London, NW9 5HT, United Kingdom

Received 9 April 2001/Returned for modification 30 May 2001/Accepted 8 July 2001

Biprobe identification assays based on real-time PCR were designed for 15 species of coagulase-negative staphylococci (CNS). Three sets of primers and four biprobes were designed from two variable regions of the 16S rRNA gene. An identification scheme was developed based on the pattern of melting peaks observed with the four biprobes that had been tested on 24 type strains. This scheme was then tested on 100 previously identified clinical isolates and 42 blindly tested isolates. For 125 of the 142 clinical isolates there was a perfect correlation between the biprobe identification and the result of the ID 32 Staph phenotypic tests and PCR. For 12 of the other isolates a 300-bp portion of the 16S rRNA gene was sequenced to determine identity. The remaining five isolates could not be fully identified. LightCycler real-time PCR allowed rapid and accurate identification of the important CNS implicated in infection.

* Corresponding author. Mailing address: Molecular Biology Unit, Virus Reference Division, Central Public Health Laboratory, 61 Colindale Ave., Colindale, London, NW9 5HT, United Kingdom. Phone: 020 8200 4400, ext. 3070. Fax: 020 8200 1569. E-mail: nsaunders{at}phls.org.uk.

Journal of Clinical Microbiology, September 2001, p. 3047-3051, Vol. 39, No. 9
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.9.3047-3051.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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