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首页> 外文期刊>Journal of Clinical Microbiology >Rapid fingerprinting of Helicobacter pylori by polymerase chain reaction and restriction fragment length polymorphism analysis.
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Rapid fingerprinting of Helicobacter pylori by polymerase chain reaction and restriction fragment length polymorphism analysis.

机译：通过聚合酶链反应和限制性片段长度多态性分析快速鉴定幽门螺杆菌。

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A simple and reliable technique was developed for differentiating Helicobacter pylori strains by restriction fragment length polymorphism analysis of polymerase chain reaction (PCR)-amplified DNAs. Oligonucleotide primer pairs developed to the urease, 48-kDa stress protein (htrA), and 26-kDa antigen-encoding genes were used to amplify fragments of the appropriate size from crude boiled cell preparations. The PCR-amplified products were digested with Sau3A, HaeIII, MspI, AluI, MluI, HinfI, and XbaI restriction endonucleases. Restriction fragment length polymorphisms were particularly evident within the urease and htrA genes and were easily detected by Sau3A, HaeIII, MspI, and AluI restriction endonuclease analysis. Double digestion of these separately amplified products or restriction analysis of multiple PCR-amplified fragments was found to discriminate 17 of 17 (100%) H. pylori strains which had unique genomic DNA fingerprints. Results of an investigation of multiple isolate sets obtained from patients before and after therapy was consistent with the hypothesis that treatment failures were due to the persistence of the same strain but did not discount the possibility that the patients were reinfected with a strain shared by family members or close contacts. The results indicate that the PCR-restriction endonuclease analysis method can be applied directly to biopsy samples, has the potential to fingerprint H. pylori isolates rapidly, and may permit detailed epidemiological investigations on the transmission of this important pathogen.
机译：通过对聚合酶链反应（PCR）扩增的DNA进行限制性片段长度多态性分析，开发了一种简单而可靠的技术来区分幽门螺杆菌。已开发出针对尿素酶，48 kDa应激蛋白（htrA）和26 kDa抗原编码基因的寡核苷酸引物对，用于从粗煮细胞制备物中扩增适当大小的片段。用Sau3A，HaeIII，MspI，AluI，MluI，HinfI和XbaI限制性核酸内切酶消化PCR扩增的产物。限制性片段长度多态性在脲酶和htrA基因中特别明显，并且可以通过Sau3A，HaeIII，MspI和AluI限制性核酸内切酶分析轻松检测到。发现对这些单独扩增的产物进行双消化或对多个PCR扩增片段进行限制性分析可区分出17个（100％）幽门螺杆菌菌株中的17个，这些菌株具有独特的基因组DNA指纹。对治疗前后从患者身上获得的多种分离株的调查结果与以下假设相符：治疗失败是由于同一菌株的持续存在，但并未降低患者再次感染家人共享的菌株的可能性。或关闭联系人。结果表明，PCR限制性核酸内切酶分析方法可直接应用于活检样本，具有快速识别幽门螺杆菌分离物的潜力，并可允许对该重要病原体的传播进行详细的流行病学研究。

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