Sachse Konrad, Ruettger Anke
Institute of Molecular Pathogenesis at Friedrich-Loeffler-Institut (Federal Research Institute for Animal Health), Naumburger Str. 96a, 07743, Jena, Germany,
Methods Mol Biol. 2015;1247:391-400. doi: 10.1007/978-1-4939-2004-4_28.
Pathogenic Chlamydia (C.) psittaci and C. trachomatis strains can be genotyped based on variations in the ompA genomic locus. In the present chapter, we describe rapid genotyping assays for both chlamydial agents using the ArrayStrip™ (AS) microarray platform. The test is targeting multiple discriminatory sites in the variable domains of the ompA gene by using 35 (C. psittaci) and 61 (C. trachomatis) oligonucleotide probes representing genotype-specific polymorphisms. In addition to discrimination among the established genotypes, this approach allows identification of atypical strains that were not accessible to typing using previously established techniques, such as PCR-RFLP or serotyping. The present DNA microarray assay can be conducted directly on clinical tissue samples and is suitable for tracing epidemiological chains and exploring the dissemination of particular genotypes. The procedure is easy to handle and economically affordable, and it allows genotyping of up to 32 clinical samples per day, thus lending itself for routine diagnosis as well.
致病性鹦鹉热衣原体和沙眼衣原体菌株可根据ompA基因组位点的变异进行基因分型。在本章中，我们描述了使用ArrayStrip™（AS）微阵列平台对这两种衣原体病原体进行快速基因分型检测的方法。该检测通过使用35个（鹦鹉热衣原体）和61个（沙眼衣原体）代表基因型特异性多态性的寡核苷酸探针，针对ompA基因可变区的多个鉴别位点。除了区分已确定的基因型外，这种方法还能鉴定出使用先前建立的技术（如PCR-RFLP或血清分型）无法分型的非典型菌株。目前的DNA微阵列检测可直接在临床组织样本上进行，适用于追踪流行病学链条和探索特定基因型的传播情况。该方法操作简便且经济实惠，每天可对多达32个临床样本进行基因分型，因此也适用于常规诊断。
