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Journal of Clinical Microbiology, August 2003, p. 3526-3531, Vol. 41, No. 8
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.8.3526-3531.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Single-Nucleotide-Polymorphism Mapping of the Pseudomonas aeruginosa Type III Secretion Toxins for Development of a Diagnostic Multiplex PCR System

Temitayo Ajayi,1,2 Leonard R. Allmond,3 Teiji Sawa,3 and Jeanine P. Wiener-Kronish1,2,4*

Department of Anesthesia and Perioperative Care,3 Cardiovascular Research Institute,1 Division of Pulmonary and Critical Care Medicine,2 Department of Medicine, University of California, San Francisco, San Francisco, California 941434

Received 4 February 2003/ Returned for modification 22 April 2003/ Accepted 1 May 2003

We mapped the coding single nucleotide polymorphisms in four toxin genes—exoS, exoT, exoU, and exoY—of the Pseudomonas aeruginosa type III secretion system among several clinical isolates. We then used this information to design a multiplex PCR assay based on the simultaneous amplification of fragments of these genes. Eight strains of known genotype were used to test our multiplex PCR method, which showed 100% sensitivity and specificity in this small sample size. This assay appears to be promising for the rapid and accurate genotyping of the presence of these genes in clinical strains of P. aeruginosa.

* Corresponding author. Mailing address: Anesthesia Department, 513 Parnassus Ave., S-261, University of California, San Francisco, CA 94143-0542. Phone: (415) 476-8968. Fax: (415) 476-8841. E-mail: WienerKJ{at}anesthesia.ucsf.edu.

Journal of Clinical Microbiology, August 2003, p. 3526-3531, Vol. 41, No. 8
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.8.3526-3531.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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