中国媒介生物学及控制杂志 ›› 2009, Vol. 20 ›› Issue (5): 460-463.

• 论著 • 上一篇    下一篇

浙江省钩端螺旋体FlaB-PCR检测方法的建立与应用

姜理平孟真陆群英罗芸叶菊莲张政   

  1. 浙江省疾病预防控制中心微生物所(杭州 310051)
  • 收稿日期:2009-06-22 出版日期:2009-10-20 发布日期:2009-10-20
  • 作者简介:姜理平(1964-),男,副主任技师,从事自然疫源性疾病检验研究。

Establishment of FlaB?PCR method used for leptospira detection and its application in Zhejiang province

 JIANG Li-Ping, MENG Zhen, LIU Qun-Ying, LUO Yun, YE Ju-Lian, ZHANG Zheng   

  1. Zhejiang Center for Disease Control and Prevention, Hangzhou 310051, Zhejiang Province, China
  • Received:2009-06-22 Online:2009-10-20 Published:2009-10-20

摘要:

  【摘要】 目的 探讨FlaB?PCR检测方法对钩端螺旋体(钩体)检测的可行性。方法 根据鞭毛蛋白B基因(FlaB)选择引物,与卫生行业标准推荐的引物G1/G2对比,进行特异性与灵敏度研究,并应用于龙游、常山两地的蛙、鼠标本钩体PCR检测。对浙江省2007年分离菌株进行钩体PCR鉴定。结果 FlaB?PCR具有较高的灵敏度和特异性,该引物可特异性扩增致病性钩体DNA,对本实验所用的其他细菌不扩增。2007年分离的钩体菌株进行PCR鉴定结果与血清学鉴定结果相符。鼠和蛙肾标本进行FlaB?PCR检测,阳性率20.94%,与卫生行业标准推荐的引物G1/G2对比,两法符合率为 90.54%。结论 FlaB?PCR检测方法可灵敏、特异地检测致病性钩体,能正确有效地反映野生动物带病毒率,可为控制钩体病提供依据。

关键词: 鞭毛蛋白B基因, 钩端螺旋体, 聚合酶链反应

Abstract:

  【Abstract】 Objective To approach the feasibility of FlaB?PCR used for the detection of leptospira. Methods The proper  primer  was  synthesized  according  to  FlaB  gene and its sensitivity and specificity was evaluated compared to the primer G1/G2 recommended by public health industry standard. It was applied to leptospira detection of frog and rats which were from Longyou and Changshan. And it was identified by PCR. Results FlaB?PCR was highly sensitive and specific, and pathogenic leptospira DNA could be specifically amplified with the selected primer. The PCR results on leptospira isolated from Zhejiang province  in  2007 were in accordance with the serology results. The positive rate of the kidneys of rats and frogs was 20.94% by FlaB?PCR. Compared with the primer G1/G2 recommended by public health industry, the coincidence of two methods was 90.54%. Conclusion FlaB?PCR can detect the pathogenicity leptospira specifically and quickly, which can reflect the carrying rate of leptospira in wild animals accurately and efficiently and provide the evidence for leptospira control.

Key words: FlaB gene, Leptospira, Polymerase chain reaction

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