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首页> 外文会议>Lancefield International Symposium on Streptococci and Streptococcal Diseases >Analysis of DNA corresponding to group A streptococcal A- and B-regions of M protein by restriction fragment length polymorphism
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Analysis of DNA corresponding to group A streptococcal A- and B-regions of M protein by restriction fragment length polymorphism

机译：用限制性片段长度多态性分析对应于组蛋白的链球菌A和B区的DNA

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摘要

The emm gene amplicons encoding the N- and partial C-terminal regions of the M protein from Group A streptococcus (GAS) isolates were amplified by PCR and digested with a restriction endonuclease, Alul. Digested PCR products were then resolved by polyacrylamide gel electropho-resis. Using one pair of primers, 13 known GAS M types showed one to four bands of PCR products. Digested with Alul, they exhibited different restriction RFLP patterns. Of 106 GAS isolates examined from normal population and patients with GAS-associated complications, 95 isolates showed RFLP patterns that corresponded to the 13 known M types. The other 11 isolates demonstrated different RFLP patterns other than the known M types. They were then analyzed by DNA sequencing and other six M types were obtained. In addition, M93 GAS was the most common from the population studied, which is consistent with a previous study of Thai GAS isolates. This PCR-RFLP protocol has the potential for the rapid screening of GAS emm types, and would be able to characterize the number of C-region in different types of GAS. This method is considerably advantageous as an alternative M typing approach for epidemiological studies in developing countries in which GAS is endemic and DNA sequencer and budgets are limited.
机译：通过PCR扩增编码来自组链球菌（气体）分离株的M蛋白的N-和部分C末端区域的EMM基因扩增子并用限制性内切核酸酶消化，ALUL。然后通过聚丙烯酰胺凝胶电泳 - Resis解析消化的PCR产物。使用一对引物，13种已知的气体M型显示出一至四带PCR产物。用Alul消化，它们表现出不同的限制RFLP模式。在正常人群中检查的106个天然气分离物，95个分离物显示与13种已知的M种类型的RFLP模式。除了已知的M类型之外，另外11个分离物示出了除此之外的不同RFLP模式。然后通过DNA测序分析它们，并获得其他六种六种类型。此外，M93气是所研究的人群中最常见的，这与对泰国天然气分离株的先前研究一致。该PCR-RFLP协议具有对气体EMM类型的快速筛选的可能性，并且能够表征不同类型气体的C区的数量。该方法显着有利地作为发展中国家流行病学研究的替代M个分类方法，其中天然气流行和DNA测序仪和预算有限。

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