Original Research ARTICLE

Front. Cell. Infect. Microbiol., 17 May 2012 | doi: 10.3389/fcimb.2012.00068

Genotyping markers used for multi locus VNTR analysis with ompA (MLVA-ompA) and multi sequence typing (MST) retain stability in Chlamydia trachomatis

Clare Labiran1, Ian N. Clarke1, Lesley T. Cutcliffe1, Yibing Wang1, Rachel J. Skilton1, Kenneth Persson2, Carina Bjartling3, Björn Herrmann4, Linus Christerson4 and Peter Marsh5*
  • 1Molecular Microbiology and Infection, School of Medicine, University of Southampton, Southampton, Hampshire, UK
  • 2Department of Clinical Microbiology, Malmö University Hospital, Malmö, Sweden
  • 3Department of Obstetrics and Gynaecology, Malmö University Hospital, Malmö, Sweden
  • 4Department of Medical Sciences, Section of Clinical Bacteriology, Uppsala University, Uppsala, Sweden
  • 5Health Protection Agency, Microbiology Services, Public Health Laboratory Southampton, Southampton General Hospital, Southampton, Hampshire, UK

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.

Keywords: Chlamydia trachomatis, MLVA-ompA, multi sequence typing, marker stability, tissue culture

Citation: Labiran C, Clarke IN, Cutcliffe LT, Wang Y, Skilton RJ, Persson K, Bjartling C, Herrmann B, Christerson L and Marsh P (2012) Genotyping markers used for multi locus VNTR analysis with ompA (MLVA-ompA) and multi sequence typing (MST) retain stability in Chlamydia trachomatis. Front. Cell. Inf. Microbio. 2:68. doi: 10.3389/fcimb.2012.00068

Received: 20 January 2012; Accepted: 30 April 2012;
Published online: 17 May 2012.

Edited by:

Yasuko Rikihisa, Ohio State University, USA

Reviewed by:

Ted Hackstadt, Rocky Mountain Laboratories/NIAID/NIH, USA
Yajun Song, Beijing Institute of Microbiology and Epidemiology, China
João P. Gomes, National Institute of Health, Portugal

Copyright: © 2012 Labiran, Clarke, Cutcliffe, Wang, Skilton, Persson, Bjartling, Herrmann, Christerson and Marsh. This is an open-access article distributed under the terms of the Creative Commons Attribution Non Commercial License, which permits non-commercial use, distribution, and reproduction in other forums, provided the original authors and source are credited.

*Correspondence: Peter Marsh, Health Protection Agency, Microbiology Services, Public Health Laboratory Southampton, Southampton General Hospital, Tremona Road, Southampton, SO16 6YD, Hampshire, UK. e-mail: peter.marsh@uhs.nhs.uk

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