Department of Microbiology and research laboratory "Microorganismes et Pathologies Humaines", Habib Bourguiba university hospital, Medical School of Sfax, University of Sfax, Sfax, Tunisia.
Biopesticides Laboratory, Centre of Biotechnology of Sfax, University of Sfax, Sfax, Tunisia.
Microbiologyopen. 2018 Apr;7(2):e00549. doi: 10.1002/mbo3.549. Epub 2017 Dec 28.
Two techniques based on ompA amplification of Chlamydia trachomatis were compared, being reverse hybridization (RHM) and ompA sequencing (OSA), to investigate the concordance between them and to study the epidemiological relevance of each method. In addition, phylogenetic analysis was performed on the ompA sequences. One hundred and seven C. trachomatis positive samples from Tunisian patients and female sex workers were analyzed using both the RHM and ompA sequencing. The overall genovar distribution obtained with both techniques was very similar. The RHM identified nine genovars, being B, D, E, F, G, H, I, J and K, where B, I, J, and K were only found in mixed infections versus 7 types for the OSA being D, E, F, G, H, I, and K. The agreement between both typing techniques was 87.8%. Both methods showed that genovar E was the most predominant type. In 24.3% of the analyzed samples, mixed infections were detected. In 96.1% of these, the genovar identified by OSA was also detected using the RHM. OmpA sequencing allowed determination of six genovar types that could not be typed using RHM. The analyses of ompA nucleotide variation in the 107 clinical specimens detected ompA genovar variants with distinct ompA mutational patterns for types D2, G1, G2, and H1. In conclusion, RHM and OSA showed a high agreement in C. trachomatis genotyping results with each having their specific benefits.
两种基于沙眼衣原体 ompA 扩增的技术，即反向杂交（RHM）和 ompA 测序（OSA），被用于比较，以调查它们之间的一致性，并研究每种方法的流行病学相关性。此外，还对 ompA 序列进行了系统发育分析。用 RHM 和 ompA 测序法分析了 107 例来自突尼斯患者和女性性工作者的沙眼衣原体阳性样本。两种技术获得的总体基因型分布非常相似。RHM 鉴定出 9 种基因型，分别为 B、D、E、F、G、H、I、J 和 K，其中 B、I、J 和 K 仅在混合感染中发现，而 OSA 仅发现 7 种基因型，分别为 D、E、F、G、H、I 和 K。两种分型技术的一致性为 87.8%。两种方法均表明基因型 E 是最主要的类型。在分析的样本中，24.3%检测到混合感染。在这些样本中，96.1%的样本使用 OSA 鉴定的基因型也可以使用 RHM 鉴定。ompA 测序可以确定 6 种不能用 RHM 分型的基因型类型。对 107 例临床标本 ompA 核苷酸变异的分析检测到了 ompA 基因型变体，其 ompA 突变模式与 D2、G1、G2 和 H1 型不同。总之，RHM 和 OSA 在沙眼衣原体基因分型结果上具有高度一致性，每种方法都有其特定的优势。
