牛结核病两种活体诊断方法检测结合病原学诊断的研究

Study on Two Technologies of Diagnosis in Vivo for Cattle Tuberculosis Combining with Etiological Detection

本文报道应用两种活体诊断方法检测牛结核病的试验结果。2010～2011年度,应用PPD皮内变态反应方法分3批次对5156头奶牛和奶水牛实施监测,监测阳性数57头,阳性率1.11%。采集该57头牛抗凝全血,应用γ-干扰素酶联免疫吸附试验进行检测,检出阳性样品3份,阳性率5.26%。对14头第一批PPD皮内变态反应阳性牛进行病理检验和进行细菌分离培养和PCR检测,结果3份样品分离培养呈阳性,其中1份PCR鉴定为结核分枝杆菌,2份为非分枝杆菌。PCR方法检测的14份组织样品中,2份为结核分枝杆菌阳性,1份为牛结核分枝杆菌阳性。结合病理剖检和病原PCR诊断,分析比较PPD皮内变态反应试验和γ-干扰素酶联免疫吸附试验的敏感性和特异性,由于PPD纯度、非特异性反应、结果判定的主观性等因素,导致PPD皮内变态反应监测检出假阳性率较高。

This paper reports the result of test for Bovine Tuberculosis by two techniques in live cattle.Total three batches of dairy cattle and dairy water buffalo including 5156 heads were tested by PPD Intradermal Allergy during a period of 2010-2011,with 57 heads positive taking up a positive rate of 1.11% in tested animal.Total 57 blood samples were collected by using anticoagulant of heparin from each of the above positive cattle,and tested by γ-interferon ELISA with 3 positive taking up a positive rate of 5.26% in tested samples.Autopsy was carried out in 14 heads among the positive cattle in the first tested batch,and 14 tissue samples were collected each of the autopsied cattle for further isolation and culture of bacteria,and confirmation by PCR method.Result showed that 3 samples of 14 could be confirmed to be positive in bacteria culture,in which 1 confirmed to Mycobacterium bovis,the other 2 are not Mycobacterium.PCR result showed that 2 samples of 14 could be confirmed to be Mycobacterium,1 samples of 14 could be confirmed to be Mycobacterium bovis.The sensitivity and specificity of the PPD test and γ-interferon ELISA are analysised primarily,combining with PCR test and Pathological profile inspection.The false positive high rate of the PPD test due to the fineness of PPD,nonspecific reaction,the Subjectivity of the result judge etc.