Open AccessGenotyping of C hlamydia trachomatis directly from urogenital and conjunctiva samples using an ompA gene pyrosequencing‐based assay
Author(s) -
Kese Darja,
Potocnik Marko,
Maticic Mojca,
Kogoj Rok
Publication year - 2011
Publication title -
fems immunology & medical microbiology
Language(s) - English
Resource type - Journals
eISSN - 1574-695X
pISSN - 0928-8244
DOI - 10.1111/j.1574-695x.2011.00843.x
Subject(s) - pyrosequencing , chlamydia trachomatis , biology , genotype , genotyping , typing , chlamydiaceae , conjunctiva , population , microbiology and biotechnology , virology , genetics , gene , immunology , medicine , environmental health
The aim of the present study was to develop and validate a multitarget pyrosequencing‐based protocol for basic C hlamydia trachomatis genotyping directly from clinical samples and to characterize the distribution of genotypes among S lovenian sexually active population. The newly developed combination of assays that targets the variable domains VD ‐I and VD ‐ IV of the C . trachomatis ompA gene, was optimized and validated with 11 reference C . trachomatis strains and by comparison to complete ompA conventional sequencing. In addition, 183 clinical specimens which were previously diagnosed as C . trachomatis positive were evaluated by pyrosequencing. The pyrosequencing products showed a 100% match to corresponding sections of the respective conventional ompA sequences. Based on our results the most frequent genotype in urogenital samples was E (51.1%) followed by F (21.4%), G and K (6.9%), D (6.1%), H (3.8%), J (2.3%) and Ia and Ja (0.8%). In conjunctiva samples the genotype distribution was E (63.3%), D and F (13.3%), K (6.7%) and G (3.3%). Pyrosequencing thus proved itself to be a rapid method for C . trachomatis typing, which is important for better understanding the pathogenesis and epidemiology of this pathogen.