Mérien F, Amouriaux P, Perolat P, Baranton G, Saint Girons I
Unité de Bactériologie Moléculaire et Médicale, Institut Pasteur, Paris, France.
J Clin Microbiol. 1992 Sep;30(9):2219-24. doi: 10.1128/jcm.30.9.2219-2224.1992.
A sensitive assay for Leptospira spp., the causative agent of leptospirosis, was developed on the basis of the polymerase chain reaction (PCR). A 331-bp sequence from the Leptospira interrogans serovar canicola rrs (16S) gene was amplified, and the PCR products were analyzed by DNA-DNA hybridization by using a 289-bp fragment internal to the amplified DNA. Specific PCR products also were obtained with DNA from the closely related nonpathogenic Leptospira biflexa but not with DNA from other spirochetes, such as Borrelia burgdorferi, Borrelia hermsii, Treponema denticola, Treponema pallidum, Spirochaeta aurantia, or more distant organisms such as Escherichia coli, Staphylococcus aureus, Mycobacterium tuberculosis, and Proteus mirabilis. The assay was able to detect as few as 10 bacteria. Leptospira DNA was detected in urine from experimentally infected mice. In addition, the test was found to be suitable for diagnosing leptospirosis in humans. Cerebrospinal fluid and urine from patients with leptospirosis were positive, whereas samples from control uninfected patients were negative.
基于聚合酶链反应（PCR）开发了一种检测钩端螺旋体病病原体钩端螺旋体属的灵敏检测方法。扩增了问号钩端螺旋体犬血清型rrs（16S）基因的一段331 bp序列，并使用扩增DNA内部的一段289 bp片段通过DNA-DNA杂交分析PCR产物。从密切相关的非致病性双曲钩端螺旋体的DNA中也获得了特异性PCR产物，但从其他螺旋体如伯氏疏螺旋体、赫氏疏螺旋体、齿垢密螺旋体、梅毒螺旋体、金色螺旋体，或更远缘的生物体如大肠杆菌、金黄色葡萄球菌、结核分枝杆菌和奇异变形杆菌的DNA中未获得。该检测方法能够检测低至10个细菌。在实验感染小鼠的尿液中检测到了钩端螺旋体DNA。此外，发现该检测方法适用于诊断人类钩端螺旋体病。钩端螺旋体病患者的脑脊液和尿液呈阳性，而未感染对照患者的样本呈阴性。
