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Journal of Clinical Microbiology, July 2004, p. 2935-2939, Vol. 42, No. 7
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.7.2935-2939.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Combination of PCR Targeting the VD2 of omp1 and Reverse Line Blot Analysis for Typing of Urogenital Chlamydia trachomatis Serovars in Cervical Scrape Specimens

Department of Pathology, Vrije Universiteit Medical Center, Amsterdam,¹ Labs for Pathology and Medical Microbiology, PAMM Institute, Eindhoven, The Netherlands,³ División Epidemiología, Instituto Nacional de Cancerología, Bogota, Colombia²

Received 9 December 2003/ Returned for modification 26 January 2004/ Accepted 4 April 2004

In this study we developed and evaluated a new PCR-based typing assay, directed to the VD2 region of the omp1 gene, for the detection and typing of urogenital Chlamydia trachomatis infections. A nested VD2 PCR-reverse line blot (RLB) assay was developed for the typing of nine different urogenital serovars of C. trachomatis. The assay developed was tested with reference strains of C. trachomatis serovars and cervical scrapes of 86 Colombian women previously found to be positive for C. trachomatis by using plasmid PCR. Two sets of primers directed to the VD2 region of the omp1 gene of C. trachomatis were designed, and fragments of 220 and 166 bp were generated in the primary and nested PCRs, respectively. In addition, an RLB assay was developed to identify nine different urogenital serovars of C. trachomatis (Ba, D, E, F, G, H, I, J, and K) and group controls, including group B (Ba, D, and E), group C (I, J, K, and H), and an intermediate group (F and G). Using this assay, we were able to type 81 of the 86 samples (94.2%). Of these samples, 91.3% were single C. trachomatis infections, and 8.7% were multiple infections. The most common serovars identified were serovars D (22.2%), F (18.5%), G (13.6%), and E (12.3%). Of the women with multiple C. trachomatis infections, >50% contained both serovars D and E. The nested VD2 PCR-RLB developed is a simple, fast, and specific method for the identification of individual urogenital C. trachomatis serovars previously detected by using plasmid PCR. Moreover, it is an appropriate method for studying multiple C. trachomatis infections and for use in large epidemiological studies.

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