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首页> 外文期刊>Journal of Clinical Microbiology >Differentiation of Helicobacter pylori strains directly from gastric biopsy specimens by PCR-based restriction fragment length polymorphism analysis without culture.
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Differentiation of Helicobacter pylori strains directly from gastric biopsy specimens by PCR-based restriction fragment length polymorphism analysis without culture.

机译：通过基于PCR的限制性片段长度多态性分析无需培养即可直接从胃活检标本中区分幽门螺杆菌菌株。

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Recent studies have shown the usefulness of PCR-based restriction fragment length polymorphism (RFLP) analysis for differentiating Helicobacter pylori strains isolated by culture. For this study, a PCR-based RFLP assay was developed for directly typing H. pylori strains from gastric biopsy specimens. Nineteen gastric biopsy specimens obtained from patients undergoing endoscopy for gastrointestinal complaints were cultured for isolation of H. pylori. Genomic DNA preparations from these gastric biopsy specimens and the corresponding H. pylori isolates were tested by our PCR-based RFLP assay. The 1,179-bp H. pylori DNA fragments amplified by the PCR assay were digested with the restriction enzymes HhaI, MboI, and AluI and analyzed by agarose gel electrophoresis. HhaI, MboI, and AluI digestion produced 11, 10, and 6 distinguishable digestion patterns, respectively, from the 19 H. pylori isolates tested and generated 13, 11, and 6 different patterns, respectively, from the 19 gastric biopsy specimens. The patterns from 13 of the 19 gastric biopsy specimens matched those of the H. pylori isolates from the corresponding patients. The patterns from the remaining six biopsy specimens appeared to represent infection by two strains of H. pylori; the pattern of one strain was identical to that of the isolate from the corresponding patient. By combining all the restriction enzyme digestion patterns obtained by using HhaI, MboI, and AluI, we observed 19 distinct RFLP patterns from the 19 specimens. The results suggest that the PCR-based RFLP analysis method may be useful as a primary technique to identify and distinguish H. pylori strains directly from gastric biopsy specimens without culture of the organisms.
机译：最近的研究表明基于PCR的限制性片段长度多态性（RFLP）分析对于区分培养分离的幽门螺杆菌菌株的有用性。对于这项研究，开发了一种基于PCR的RFLP分析方法，用于直接从胃活检标本中分离幽门螺杆菌菌株。培养十九个胃镜检查样本，取自接受内镜检查的胃肠道不适患者，以分离幽门螺杆菌。通过基于PCR的RFLP分析，对这些胃活检标本和相应的幽门螺杆菌分离物的基因组DNA制剂进行了测试。通过PCR分析扩增的1,179 bp幽门螺杆菌DNA片段用限制酶HhaI，MboI和AluI消化，并通过琼脂糖凝胶电泳进行分析。 HhaI，MboI和AluI消化分别从测试的19种幽门螺杆菌分离物中产生11种，10种和6种不同的消化模式，并从19种胃活检样品中分别产生13种，11种和6种不同的消化模式。 19个胃活检标本中有13个的模式与相应患者的幽门螺杆菌分离株相匹配。其余六个活检标本的模式似乎代表了两种幽门螺杆菌的感染。一种菌株的模式与相应患者分离株的模式相同。通过组合使用HhaI，MboI和AluI获得的所有限制酶消化图谱，我们从19个标本中观察到19个不同的RFLP图谱。结果表明，基于PCR的RFLP分析方法可用作直接从胃活检标本中鉴定和区分幽门螺杆菌菌株的主要技术，而无需进行生物体培养。

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