乳牛布鲁氏菌病病原DNA快速检测技术的研究

Study on rapid detection technique of DNA in Brucella abortus from dairy cows with brucellosis

在国内首次建立了乳牛原乳中布鲁氏菌外膜蛋白(OMP)25 ku基因套式聚合酶链反应(nested-PCR)检测技术。结果显示,本方法的特异性好,只能扩增布鲁氏菌DNA,而不能扩增同样能引起乳牛流产的鹦鹉热衣原体、弓形虫、胎儿弯杆菌DNA;乳样的检测灵敏度达50 CFU/mL,重复试验35次,可重复性和稳定性均十分理想;对 4 批 331 头凝集试验阳性(24 份)或阴性(307 份)乳牛的乳样用本方法检测,符合率为100%。在48 h内即可获得诊断结果。本方法同样可用于血液或其他流产病料中布鲁氏菌DNA的检测。

The nested-PCR for detecting the OMP 25ku gene of Brucella abortus in the raw milk from dairy cows was developed. The method can detect only Brucella spp and neither Chlamydia psittaci, (Toxoplasma) gondii nor Campylobacter fetus were crossreacted. The sensitivity of this method was 50(CFU/mL) milk sample and its stabilityand reproducibility were also proved. The milk samples from 24 dairy cows positive for brucellosis with the serum agglutination test were examined by the method and all of the milk specimens were positive and the diagnostic result can be obtained within 48 hours. The method can (also) be used for the examination of Brucella DNA in serum or other aborted pathologic materials.