首页> 美国卫生研究院文献>Applied and Environmental Microbiology >Effectiveness of Enterobacterial Repetitive Intergenic Consensus PCR and Random Amplified Polymorphic DNA Fingerprinting for Helicobacter pylori Strain Differentiation
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Effectiveness of Enterobacterial Repetitive Intergenic Consensus PCR and Random Amplified Polymorphic DNA Fingerprinting for Helicobacter pylori Strain Differentiation

机译:肠杆菌重复基因间共识PCR和随机扩增多态性DNA指纹图谱对幽门螺杆菌菌株分化的有效性。

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摘要

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:我们比较了肠道细菌重复基因间共识(ERIC)和随机扩增多态性DNA(RAPD)指纹图谱方法在秘鲁棚户区居民中检测到混合幽门螺杆菌感染病例的鲁棒性和区分力。培养了来自63名参与者的幽门螺杆菌分离株,并通过ERIC-PCR和RAPD测试使用了四个10个核苷酸的引物(每个反应一个引物)分析了五个单个菌落和每个参与者的其他菌落库。 ERIC和RAPD图谱之间有94%的一致性,可将分离株分类为统一与紧密相关,但不相同与可能无关,这表明kappa统计值很高,为0.8942。相关ERIC或RAPD模式的细微差异可能反映了菌株之间的基因转移,重组和/或突变,而明显不同的模式反映了不相关菌株的感染。感染棚户区的居民中至少有一半似乎携带一种以上的幽门螺杆菌菌株,尽管在31人中有19人与这些菌株密切相关。为了达到一项ERIC测试的敏感性,需要进行三个RAPD测试,每个测试使用不同的引物。 ERIC-PCR构成了幽门螺杆菌菌株区分的资源高效且省时的方法。

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