应用套式PCR检测6种血清型钩端螺旋体

DETECTION OF SIX SEROVARS OF PATHOGENIC LEPTOSPIRES BY NESTED PCR

目的 用套式 Ｐ Ｃ Ｒ 快速检测水沟和丛林等地的钩端螺旋体。方法 选 Ｌｅｐｔｏｓｐｉｒａ ｂｏｒｇｐｅｔｅｒｓｅｎｉｉ 中的一段保守序列 Ｉ Ｓ１５３３ ， 设计两对引物， 用套式 Ｐ Ｃ Ｒ 检测六种不同血清型的钩体。结果 ６ 株致病株均出现单一２４６ｂｐ 的特异性扩增产物， 而腐生株、空白对照无任何 Ｄ Ｎ Ａ 扩增条带。用琼脂糖凝胶电泳， 能检测５００ｆｇ 模板的扩增产物。结论 套式 Ｐ Ｃ Ｒ 是一种灵敏度高、特异性强的快速检测自然疫源中钩体的有效方法。

Aim To rapidly detect Leptospira in ditch and jungle by nested PCR.Methods Two sets of primers were derived from an inserted sequence IS1533 of Leptospira borgpetersenii.Six serotypes Leptospires were detected by nested PCR.Results\ A specific amplification product with 246bp length could be found from six serovar pathogenic Leptospires by nested PCR,and agarose gel electrophoresis is demonstrated to have a sensitivity of detecting 500fg DNA amplification products.Conclusion\ Nested PCR is a rapid,specific and sensitive method for detecting Leptospira in the epidemic regions.