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用于检测两种人类支原体物种(穿透支原体和人型支原体)的基于聚合酶链反应(PCR)的检测方法的开发。

Development of PCR-based assays for the detection of two human mollicute species, Mycoplasma penetrans and M. hominis.

作者信息

Grau O, Kovacic R, Griffais R, Launay V, Montagnier L

机构信息

Unité d'Oncologie Virale, Institut Pasteur, Paris, France.

出版信息

Mol Cell Probes. 1994 Apr;8(2):139-47. doi: 10.1006/mcpr.1994.1019.

Abstract

Recently, a 16S rDNA-based polymerase chain reaction (PCR) assay was developed for the selective and sensitive detection of Mycoplasma pirum. In this study, the same procedure was used in order to selectively detect by PCR two human mycoplasmas, M. hominis and M. penetrans, with a high level of sensitivity even in a context of human DNA. For each assay, the specificity was verified by testing DNA from other mollicute species (including those closely related to the corresponding mycoplasma), from bacteria phylogenetically close to mollicutes, from Escherichia coli and from human peripheral blood mononuclear cells (PBMCs). Each assay proved to be highly sensitive since it reliably detected 10 DNA molecules, even in a context of human DNA. The results of this study demonstrate the suitability of our procedure using primers which were designed for the PCR detection of human mollicutes with a high specificity and a low and reproducible threshold of sensitivity.

摘要

最近,开发了一种基于16S rDNA的聚合酶链反应(PCR)检测方法,用于选择性和灵敏地检测梨支原体。在本研究中,采用相同的程序,通过PCR选择性检测两种人支原体,即人型支原体和解脲脲原体,即使在人DNA存在的情况下也具有高度敏感性。对于每种检测方法,通过检测来自其他柔膜菌属物种(包括与相应支原体密切相关的物种)、与柔膜菌系统发育相近的细菌、大肠杆菌和人外周血单核细胞(PBMC)的DNA来验证其特异性。每种检测方法都被证明具有高度敏感性,因为即使在人DNA存在的情况下,它也能可靠地检测到10个DNA分子。本研究结果表明,我们使用为PCR检测人柔膜菌而设计的引物的程序具有适用性,该程序具有高特异性和低且可重复的敏感性阈值。

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