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Effectiveness of Enterobacterial Repetitive Intergenic Consensus PCR and Random Amplified Polymorphic DNA Fingerprinting for Helicobacter pylori Strain Differentiation

机译:细菌重复性癌基因术共识的有效性和随机扩增多态性DNA对幽门螺杆菌应变分化的指纹识别

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We compared the robustness and discriminatory power of the enterobacterial repetitive intergenic consensus (ERIC) and random amplified polymorphic DNA (RAPD) fingerprinting methods for detecting cases of mixed Helicobacter pylori infection in Peruvian shantytown residents. H. pylori isolates from 63 participants were cultured, and five single colonies and a pool of additional colonies from each participant were analyzed by ERIC-PCR and by RAPD tests with four 10-nucleotide primers (one primer per reaction). There was 94% agreement between the ERIC and RAPD profiles in classifying sets of isolates as uniform versus closely related but not identical versus probably unrelated, indicating a high kappa statistic of 0.8942. Subtle differences in related ERIC or RAPD patterns likely reflect gene transfer between strains, recombination, and/or mutation, whereas markedly different patterns reflect infection by unrelated strains. At least half of infected shantytown residents seemed to carry more than one H. pylori strain, although in 19 of 31 persons, the strains were closely related. Three RAPD tests, each with a different primer, were needed to achieve the sensitivity of one ERIC test. ERIC-PCR constitutes a resource- and time-efficient method for H. pylori strain differentiation.
机译:我们比较了肠杆菌重复性非基因共识(ERIC)和随机扩增的多晶型DNA(RAPD)指纹识别方法的鲁棒性和歧视力,用于检测秘鲁山楼居民混合幽门螺杆菌感染病例。 H.培养来自63名参与者的幽门螺杆菌分离株,通过Eric-PCR分析了来自每个参与者的五个单个菌落和来自每个参与者的其他菌落,并通过RAPD试验用四个10核苷酸引物(每次反应一个引物)进行分析。在分类的eric和RAPD配置文件之间,将分离套件组的统一与密切相关但不相同的相比可能不相关的恰到好处,表明了0.8942的高kappa统计数据,有94 %。相关埃里克或RAPD模式的微妙差异可能反映菌株,重组和/或突变之间的基因转移,而显着不同的模式反映了不相关的菌株的感染。虽然在31人的19人中,但至少有一半的受感染的棚户区居民似乎携带了多于一个H.幽门螺杆菌菌株。需要三个RAPD测试,每个RAPD测试都需要不同的底漆,以达到一个ERIC测试的敏感性。 ERIC-PCR构成了H.幽门螺杆菌应变分化的资源和时间效率。

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