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基于小亚基核糖体RNA序列引物的聚合酶链式反应检测贝氏等孢球虫

Detection of Isospora belli by polymerase chain reaction using primers based on small-subunit ribosomal RNA sequences.

作者信息

Müller A, Bialek R, Fätkenheuer G, Salzberger B, Diehl V, Franzen C

机构信息

Department of Pediatrics, University of Cologne, Germany.

出版信息

Eur J Clin Microbiol Infect Dis. 2000 Aug;19(8):631-4. doi: 10.1007/s100960000322.

Abstract

The aim of the present study was to use small-subunit (SSU)-rRNA sequences of Isospora belli to design specific primer pairs and a hybridization probe for the detection of Isospora belli in human samples by PCR and Southern blot hybridization. PCR amplification with the primer pairs produced correct DNA fragments with target DNA from samples of Isospora belli-infected patients and from cloned SSU-rRNA of Isospora belli. The nature of the PCR products was confirmed by Southern blot hybridization. No amplification was seen with template DNA extracted from other parasites. Although Isospora belli infections can be easily diagnosed using light microscopy, molecular-based techniques may prove useful as an additional diagnostic tool.

摘要

本研究的目的是利用贝氏等孢球虫的小亚基(SSU)-rRNA序列设计特异性引物对和杂交探针,通过聚合酶链反应(PCR)和Southern印迹杂交检测人体样本中的贝氏等孢球虫。用这些引物对进行PCR扩增,从贝氏等孢球虫感染患者的样本以及贝氏等孢球虫克隆的SSU-rRNA的靶DNA中产生了正确的DNA片段。通过Southern印迹杂交证实了PCR产物的性质。从其他寄生虫提取的模板DNA未出现扩增。虽然使用光学显微镜可以很容易地诊断贝氏等孢球虫感染,但基于分子的技术可能作为一种额外的诊断工具被证明是有用的。

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