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基于编码溶血相关蛋白-1的hap1基因的致病性钩端螺旋体特异性聚合酶链反应检测法。

Polymerase chain reaction assay specific for pathogenic Leptospira based on the gene hap1 encoding the hemolysis-associated protein-1.

作者信息

Branger Christine, Blanchard Béatrice, Fillonneau Catherine, Suard Isabelle, Aviat Florence, Chevallier Bruno, André-Fontaine Geneviève

机构信息

Leptospira Medical and Molecular Bacteriology Unit, ENVN, BP 40706, 44307 Nantes Cedex 03, France.

出版信息

FEMS Microbiol Lett. 2005 Feb 15;243(2):437-45. doi: 10.1016/j.femsle.2005.01.007.

Abstract

In this study, we used Southern hybridization of genomic DNA with the integral hap1 gene as a probe to show that this gene is only present in pathogenic Leptospira strains. We then selected PCR primers based on the hap1 gene, and tested them on several Leptospira strains and biological samples. Specific amplification was obtained for all pathogenic strains tested. Negative PCR results were observed with all saprophytic leptospire strains used as well as with other spirochetes and bacteria commonly found in biological samples. The results of direct PCR performed on biological samples, such as blood, urine or kidneys correlated with the results obtained with the classical Leptospira tests (culture and MAT). A PCR assay based on this gene would be a very useful tool for the rapid, sensitive and specific identification of pathogenic leptospires in samples for diagnosis or epidemiological survey.

摘要

在本研究中,我们使用基因组DNA与完整的hap1基因进行Southern杂交,以表明该基因仅存在于致病性钩端螺旋体菌株中。然后,我们基于hap1基因选择了PCR引物,并在几种钩端螺旋体菌株和生物样品上进行了测试。对所有测试的致病菌株均获得了特异性扩增。以所有用作对照的腐生钩端螺旋体菌株以及生物样品中常见的其他螺旋体和细菌进行PCR检测,结果均为阴性。对血液、尿液或肾脏等生物样品进行直接PCR的结果与经典钩端螺旋体检测(培养和显微镜凝集试验)的结果相关。基于该基因的PCR检测将是用于在诊断或流行病学调查样品中快速、灵敏和特异性鉴定致病性钩端螺旋体的非常有用的工具。

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