title: Evaluation of high-resolution typing methods for Chlamydia trachomatis in samples from heterosexual couples.
abstract: We aimed to compare conventional ompA typing of Chlamydia trachomatis with multilocus sequence typing (MLST) and multilocus variable-number tandem-repeat (VNTR) analysis (MLVA). Previously used MLST and MLVA systems were compared to modified versions that used shorter target regions and nested PCR. Heterosexual couples were selected from among persons with urogenital C. trachomatis infections visiting the sexually transmitted infection outpatient clinic in Amsterdam, The Netherlands. We identified 30 couples with a total of 65 C. trachomatis-positive samples on which MLST and MLVA for eight target regions were performed. All regions were successfully sequenced in 52 samples, resulting in a complete profile for 18 couples and 12 individuals. Nine ompA genovars from D to K, with two variants of genovar G, were found. The numbers of sequence type and MLVA type profiles were 20 for MLST and 21 for MLVA, and a combination of MLST and MLVA yielded 28 profiles, with discriminatory indexes (D) ranging from 0.95 to 0.99. Partners in 17 couples shared identical profiles, while partners in 1 couple had completely different profiles. Three persons had infections at multiple anatomical locations, and within each of these three individuals, all profiles were identical. The discriminatory capacity of all MLST and MLVA methods is much higher than that of ompA genotyping (D = 0.78). No genotype variation was found within the samples of the same person or from heterosexual couples with a putative single transmission. This shows that the chlamydial genome in clinical specimens has an appropriate polymorphism to enable epidemiological cluster analysis using MLST and MLVA.
authors.affiliation: Public Health Laboratory, Public Health Service of Amsterdam, Nieuwe Achtergracht 100, 1018 WT Amsterdam, The Netherlands. rbom@ggd.amsterdam.nl
authors.forename: Reinier J M
authors.initials: RJ
authors.lastname: Bom
authors.forename: Linus
authors.initials: L
authors.lastname: Christerson
authors.forename: Maarten F
authors.initials: MF
authors.lastname: Schim van der Loeff
authors.forename: Roel A
authors.initials: RA
authors.lastname: Coutinho
authors.forename: Björn
authors.initials: B
authors.lastname: Herrmann
authors.forename: Sylvia M
authors.initials: SM
authors.lastname: Bruisten
mesh_terms: D000328:Adult; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics; D002692:Chlamydia trachomatis / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification; D005191:Family Characteristics; D005260:Female; D020010:Heterosexuality; D006801:Humans; D008219:Lymphogranuloma Venereum / Q000382:microbiology*; D008297:Male; D018598:Minisatellite Repeats; D058889:Molecular Typing / Q000379:methods*; D058885:Multilocus Sequence Typing; D009426:Netherlands; D016133:Polymerase Chain Reaction
chemical_list: D001425:Bacterial Outer Membrane Proteins; C078167:OMPA outer membrane proteins