应用PCR方法对三种新型实验动物钩端螺旋体感染情况的调查研究

目的：建立有效的钩端螺旋体 PCR 检测方法，并对树鼩（（tree shrew， Tupaia belangeri））、长爪沙鼠（ Meriones unguiculatus； Mongolian gerbil）和灰仓鼠（ Cricetulus migratorius）等三种新型实验动物进行感染情况调查。方法针对 NCBI 公布的钩端螺旋体序列，设计并筛选特异性引物，优化 PCR 体系，进行特异性和敏感性测试；并运用优化 PCR 方法对树鼩、长爪沙鼠和灰仓鼠样品进行检测。结果成功建立钩端螺旋体 PCR 检测方法，序列测定验证了该方法的特异性。普通级树鼩钩端螺旋体的阳性率为8.33％，普通级长爪沙鼠钩端螺旋体为100％，清洁级长爪沙鼠和清洁级灰仓鼠钩端螺旋体的阳性率为0％。结论本研究建立了钩端螺旋体 PCR 检测方法，调查了树鼩、长爪沙鼠和灰仓鼠三种新型实验动物的感染情况，为这三种实验动物的研究和使用奠定基础。

Investigation of Leptospira infection in three new experimental animals by PCR methods

Objective To establish an effective PCR assay for leptospirosis detection, and applicate the assay in tree shrew, mongolian gerbil and gray hamster. Methods Sequence of leptospira was obtained from the NCBI Genbank, and primers were designed based on the sequences. The positive amplified fragments were sequenced to verify the reliability of the method. The samples from tree shrew, mongolian gerbils and hamsters were tested using this PCR method. Results The PCR method for detection of leptospirosis was successfully established. The positive rate of Leptospira was 8.33% in 60 samples of conventional tree shrews, 100% in 104 samples of the conventional Mongolian gerbils, and 0% in 60 samples of clean gray hamsters. Conclusions The establishment of this PCR assay is useful in the detection of leptospirosis in tree shrew, mongolian gerbil and gray hamster. The results of our investigation of leptospira infection levels of the three new experimental animals may promote their application in biomedical research.