title: Combination of PCR targeting the VD2 of omp1 and reverse line blot analysis for typing of urogenital Chlamydia trachomatis serovars in cervical scrape specimens.
abstract: 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.
authors.affiliation: Department of Pathology, Vrije Universiteit Medical Center, Amsterdam, The Netherlands.
authors.forename: Monica
authors.initials: M
authors.lastname: Molano
authors.forename: Chris J L M
authors.initials: CJ
authors.lastname: Meijer
authors.forename: Servaas A
authors.initials: SA
authors.lastname: Morré
authors.forename: Rene
authors.initials: R
authors.lastname: Pol
authors.forename: Adriaan J C
authors.initials: AJ
authors.lastname: van den Brule
mesh_terms: D015373:Bacterial Typing Techniques / Q000379:methods*; D002584:Cervix Uteri / Q000382:microbiology*; D002690:Chlamydia Infections / Q000382:microbiology*; D002692:Chlamydia trachomatis / Q000145:classification*; D005260:Female; D052776:Female Urogenital Diseases / Q000382:microbiology*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D018272:Porins / Q000235:genetics*; D012680:Sensitivity and Specificity
chemical_list: D018272:Porins; C071677:omp1 protein, Chlamydia trachomatis