Smythe Lee D, Smith Ina L, Smith Greg A, Dohnt Michael F, Symonds Meegan L, Barnett Leonie J, McKay David B
WHO/FAO/OIE Collaborating Centre for Reference & Research on Leptospirosis, Centre for Public Health Sciences, Queensland Health Scientific Services, Brisbane, Australia.
BMC Infect Dis. 2002 Jul 8;2:13. doi: 10.1186/1471-2334-2-13.
Leptospirosis is an emerging infectious disease. The differential diagnosis of leptospirosis is difficult due to the varied and often "flu like" symptoms which may result in a missed or delayed diagnosis. There are over 230 known serovars in the genus Leptospira. Confirmatory serological diagnosis of leptospirosis is usually made using the microscopic agglutination test (MAT) which relies on the use of live cultures as the source of antigen, often performed using a panel of antigens representative of local serovars. Other techniques, such as the enzyme linked immunosorbent assay (ELISA) and slide agglutination test (SAT), can detect different classes of antibody but may be subject to false positive reactions and require confirmation of these results by the MAT.
The polymerase chain reaction (PCR) has been used to detect a large number of microorganisms, including those of clinical significance. The sensitivity of PCR often precludes the need for isolation and culture, thus making it ideal for the rapid detection of organisms involved in acute infections. We employed real-time (quantitative) PCR using TaqMan chemistry to detect leptospires in clinical and environmental samples.
The PCR assay can be applied to either blood or urine samples and does not rely on the isolation and culture of the organism. Capability exists for automation and high throughput testing in a clinical laboratory. It is specific for Leptospira and may discriminate pathogenic and non-pathogenic species. The limit of detection is as low as two cells.
钩端螺旋体病是一种新发传染病。由于其症状多样且常类似“流感”，钩端螺旋体病的鉴别诊断较为困难，这可能导致漏诊或诊断延迟。钩端螺旋体属已知血清型超过230种。钩端螺旋体病的确证血清学诊断通常采用显微镜凝集试验（MAT），该试验依赖使用活培养物作为抗原来源，通常使用一组代表当地血清型的抗原进行检测。其他技术，如酶联免疫吸附测定（ELISA）和玻片凝集试验（SAT），可检测不同类别的抗体，但可能会出现假阳性反应，且需要通过MAT来确认这些结果。
聚合酶链反应（PCR）已被用于检测大量微生物，包括具有临床意义的微生物。PCR的敏感性常常使得无需进行分离培养，因此它非常适合快速检测急性感染中涉及的病原体。我们采用TaqMan化学法的实时（定量）PCR来检测临床和环境样本中的钩端螺旋体。
PCR检测法可应用于血液或尿液样本，且不依赖于病原体的分离培养。临床实验室具备实现自动化和高通量检测的能力。它对钩端螺旋体具有特异性，并且可以区分致病性和非致病性菌种。检测限低至两个菌体。
