Public Health Laboratory, Public Health Service of Amsterdam, Nieuwe Achtergracht 100, 1018 WT Amsterdam, The Netherlands.
J Clin Microbiol. 2011 Aug;49(8):2844-53. doi: 10.1128/JCM.00128-11. Epub 2011 Jun 8.
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.
我们旨在比较沙眼衣原体的传统ompA 分型与多位点序列分型(MLST)和多位点可变数目串联重复分析(MLVA)。将比较以前使用的 MLST 和 MLVA 系统与使用较短目标区域和嵌套 PCR 的修改版本。从荷兰阿姆斯特丹性传播感染门诊就诊的泌尿生殖道沙眼衣原体感染患者中选择异性恋夫妇。我们确定了 30 对夫妇,共有 65 份沙眼衣原体阳性样本,对其进行了 8 个目标区域的 MLST 和 MLVA。在 52 个样本中成功地对所有区域进行了测序,从而确定了 18 对夫妇和 12 个人的完整图谱。发现了从 D 到 K 的 9 种 ompA 基因型,其中包括 2 种 G 基因型变体。MLST 的序列型和 MLVA 型谱数分别为 20 个和 21 个,而 MLST 和 MLVA 的组合产生了 28 个谱,鉴别指数(D)范围为 0.95 至 0.99。17 对夫妇的伴侣具有相同的图谱,而 1 对夫妇的伴侣具有完全不同的图谱。有 3 人在多个解剖部位感染,并且在这 3 个人中的每一个人中,所有图谱均相同。所有 MLST 和 MLVA 方法的鉴别能力均远高于 ompA 基因分型(D = 0.78)。同一人或具有假定单一传播的异性恋夫妇的样本中未发现基因型变异。这表明临床标本中的衣原体基因组具有适当的多态性,可使用 MLST 和 MLVA 进行流行病学聚类分析。