Zhang Y, Li S, Dai B
Hua Xi Yi Ke Da Xue Xue Bao. 1993 Sep;24(3):262-7.
Based upon the polymerase chain reaction (PCR), We have developed a sensitive assay for Leptospira interrogans, the agent of leptospirosis. DNA amplification was carried out using primer A: 5'GATCTAATTCGCTGTAGCAGG3' and primer B: 5'ACTTTCACCCTCTATGGTCGG3'. After 30 cycles of amplification, the product could be detected by agarose gel electrophoresis. A segment (124 bp) was amplified in all strains of L. interrogans including 20 Serogroups, 49 Serovars tested, but it was not detected in Patoc I strain, Serovar patoc of Leptospira biflexa and 3055 strain of Leptonema illini (both of which are nonpathogenic). All of the serum (first time) which proved either by blood culture or MAT showed that positive rates were 100%. Leptospiral infection in humans and some domestic animals leads to one or more of a variety of manifestation and persists through a considerable duration of time. However it is relatively difficult to demonstrate the presence of leptospires in serum. The serum (second time) obtained from 55 patients with leptospirosis (6-60 days after on set) showed that PCR positive rates were 74.55% (41/55). The PCR positive rates for healthy subjects were 15% (3/20), P < 0.001. The diagnosis of leptospirosis by using PCR may become a significant addition to the routine laboratory diagnosis and a valuable technique for the investigation of leptospirosis pathogenesis.
基于聚合酶链反应（PCR），我们开发了一种针对钩端螺旋体病病原体问号钩端螺旋体的灵敏检测方法。使用引物A：5'GATCTAATTCGCTGTAGCAGG3'和引物B：5'ACTTTCACCCTCTATGGTCGG3'进行DNA扩增。经过30个循环的扩增后，产物可通过琼脂糖凝胶电泳检测到。在所有测试的问号钩端螺旋体菌株中均扩增出一段124bp的片段，这些菌株包括20个血清群、49个血清型，但在双曲钩端螺旋体帕托克血清型帕托克I菌株和伊利尼纤毛菌3055菌株（均为非致病性）中未检测到。所有经血培养或显微镜凝集试验（MAT）证实的血清（首次）显示阳性率均为100%。人和一些家畜感染钩端螺旋体会导致多种表现中的一种或多种，并持续相当长一段时间。然而，在血清中证明钩端螺旋体的存在相对困难。从55例钩端螺旋体病患者（发病后6 - 60天）获得的血清（第二次）显示PCR阳性率为74.55%（41/55）。健康受试者的PCR阳性率为15%（3/20），P<0.001。使用PCR诊断钩端螺旋体病可能成为常规实验室诊断的重要补充，也是研究钩端螺旋体病发病机制的一项有价值的技术。
