Genotyping of Chlamydophila psittaci using a new DNA microarray assay based on sequence analysis of ompA genes.
Genotyping of Chlamydophila psittaci using a new DNA microarray assay based on sequence analysis of ompA genes.
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基于OMPA基因的序列分析，使用新的DNA微阵列测定法对衣原体dophila psittaci进行基因分型。
DOI：
10.1186/1471-2180-8-63
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发表时间：
2008-04-17
期刊：
影响因子：
4.2
通讯作者：
Slickers, Peter
中科院分区：
文献类型：
--
作者：
Sachse, Konrad;Laroucau, Karine;Hotzel, Helmut;Schubert, Evelyn;Ehricht, Ralf;Slickers, Peter
目前使用的禽源人畜共患病病原体嗜衣原体（Chlamaviophila（C.）鹦鹉热是从血清学进化而来的，并且基于ompA序列变异。它包括7种鸟类和2种非鸟类基因型。扩增的ompA基因的限制性内切酶切割和不太常见的ompA测序被用于检查，但是，除了方法上的限制外，越来越多的最近测试的菌株不能被分配到任何确定的基因型。对所有可获得的ompA基因序列的综合分析揭示了C.鹦鹉热，这只是部分涵盖了目前的基因分型方案。我们建议调整和扩展目前的计划，其中包括引入亚组更异质的基因型A，E/B和D，以及6个临时基因型代表迄今无法分型菌株。序列分析的结果已被纳入一个新的DNA微阵列的设计。基于ArrayTube™微阵列的ompA基因分型检测已被证明可以区分已建立的基因型，并识别迄今为止未分型的菌株。它的高特异性，这使得单核苷酸多态性的检测，是由于平行的方法，包括在使用来自ompA基因的可变结构域2和4的35个杂交探针。传统的基因分型系统不能充分反映C.鹦鹉新开发的基于DNA微阵列的检测代表了一种有前途的诊断工具，用于追踪流行病学链，探索基因型的传播和识别C.鹦鹉
The currently used genotyping system for the avian zoonotic pathogen Chlamydophila (C.) psittaci has evolved from serology and is based on ompA sequence variations. It includes seven avian and two non-avian genotypes. Restriction enzyme cleavage of the amplified ompA gene and, less frequently, ompA sequencing are being used for examination, but, beside methodological limitations, an increasing number of recently tested strains could not be assigned to any established genotype. Comprehensive analysis of all available ompA gene sequences has revealed a remarkable genetic diversity within the species C. psittaci, which is only partially covered by the present genotyping scheme. We suggest adjustments and extensions to the present scheme, which include the introduction of subgroups to the more heterogeneous genotypes A, E/B and D, as well as six provisional genotypes representing so far untypable strains. The findings of sequence analysis have been incorporated in the design of a new DNA microarray. The ArrayTube™ microarray-based ompA genotyping assay has been shown to discriminate among established genotypes and identify so far untyped strains. Its high specificity, which allows detection of single-nucleotide polymorphisms, is due to the parallel approach consisting in the use of 35 hybridization probes derived from variable domains 2 and 4 of the ompA gene. The traditional genotyping system does not adequately reflect the extent of intra-species heterogeneity in ompA sequences of C. psittaci. The newly developed DNA microarray-based assay represents a promising diagnostic tool for tracing epidemiological chains, exploring the dissemination of genotypes and identifying non-typical representatives of C. psittaci.
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