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Abstract
We aimed to evaluate the stability of the Chlamydia trachomatis multi locus VNTR analysis (MLVA-ompA) and multi sequence typing (MST) systems through multiple passages in tissue culture. Firstly, we analyzed the stability of these markers through adaptation of C. trachomatis to tissue culture and secondly, we examined the stability of a four-locus MLVA-ompA and a five-locus MST system after multiple passages in tissue culture. Marker sequences were monitored through successive chlamydial developmental cycles to evaluate the stability of the individual DNA markers through many bacterial divisions and this, in turn, informed us of the usefulness of using such typing systems for short and long-term molecular epidemiology. Southampton genitourinary medicine (GUM) clinic isolates from endocervical swabs collected from C. trachomatis positive women were passaged through tissue culture. MLVA-ompA typing was applied to primary swab samples and to the same samples after C. trachomatis had been passaged through cell culture (eight passages). Sequence data from time-zero and passage-eight isolates were aligned with reference sequences to determine the stability of the markers. The Swedish new variant (nvCT) underwent 72 passages in cell culture and the markers of the two schemes were similarly analyzed. Analysis of genetic markers of the MLVA-ompA typing system before and after the isolates were introduced to tissue culture showed no change in the dominant sequence. The nvCT that had been passaged 72 times over the duration of a year also showed no variation in the dominant sequence for both the genotyping schemes. MLVA-ompA and MST markers are stable upon adaptation of C. trachomatis to tissue culture following isolation of strains from primary endocervical swab samples. These markers remain stable throughout multiple rounds of cell-division in tissue culture, concomitant with the incubation period and appearance of symptoms normally associated with host-infection. Both genotyping schemes are, therefore, suitable for epidemiology of C. trachomatis.
Highlights
Chlamydia trachomatis, an intracellular bacterial pathogen is the leading cause of sexually transmitted infections (STIs) worldwide
We aimed to evaluate the stability of the Chlamydia trachomatis multi locus variable number tandem repeat (VNTR) analysis (MLVA-outer membrane protein A (ompA)) and multi sequence typing (MST) systems through multiple passages in tissue culture
All isolates selected for marker stability had to have different ompA genotypes and they all had to have gone through the same number of passages in order for the data to be comparable
Summary
An intracellular bacterial pathogen is the leading cause of sexually transmitted infections (STIs) worldwide. C. trachomatis is currently divided into at least 15 serotypes. Genital infections are caused by the non-invasive serotypes D to K and by the invasive lymphogranuloma venereum (LGV) which infects the lymphatic system. If left untreated, which is the case for up to 50% of men and 70% of women due to the asymptomatic nature of this STI, a range of complications may persist, opportunistic screening and partner notification detect a lot of asymptomatic cases. Individuals infected with LGV can progress to develop proctitis, lymphadenopathy, and genital ulcerations, whereas disease outcome for individuals infected with serotypes D to K range from infertility, ectopic pregnancy, and pelvic inflammatory disease in women and epididymitis in men
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