牛布鲁氏菌A19号疫苗免疫血清几种检测方法的比较
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S851.34

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乌鲁木齐市科技局计划项目(P161210004)


Comparison of Several Methods for Detecting Immune Serum with Bovine Brucella A19 Vaccine
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    摘要:

    为选择科学适用的牛布鲁氏菌病净化检测方法,对1 721份免疫布鲁氏菌A19号疫苗18个月后的牛血清,分别用虎红平板凝集试验(RBT)、试管凝集试验(SAT)、竞争酶联免疫吸附试验(cELISA)、间接酶联免疫吸附试验(iELISA)和荧光偏振试验(FPA)5种血清学检测方法,以及环介导等温扩增技术(LAMP)和荧光定量 PCR病原学检测方法进行检测,分析比对不同检测方法的特异性、敏感性、误诊率、漏诊率、符合率、Kappa值等。结果显示:与SAT相比,其他4种血清学检测方法的敏感性、特异性和符合率从高到低依次为iELISA、FPA、cELISA、RBT,其中敏感性均在85.00%以上,特异性均在97.60%以上,符合率均在97.50%以上,Kappa值均≥1.00;与PCR相比,LAMP方法敏感性低(9.09%),但特异性强(99.65%),与PCR符合率高(98.49%)。结果表明:血清学检测方法较为敏感特异,符合率和一致性均较高,而分子生物学检测方法特异性强,不易误诊和漏诊。因此,对于牛群的布鲁氏菌病净化,要结合牛群污染和免疫情况以及净化的不同阶段选择适用的检测方法,仅用一种方法可能会存在偏差。建议先用iELISA或FPA或RBT进行初筛,再用cELISA或SAT进行确诊,进而对确诊为阳性牛的阴道拭子或奶样进行分子生物学检测,以确定是否存在布鲁氏菌感染。本研究为免疫牛群的布鲁氏菌病净化检测方法选择提供了参考。

    Abstract:

    In order to select a scientific and applicable method to purify and detect bovine brucellosis,1 721 serums collected from the cattle vaccinated with Brucella A19 vaccine for 18 months were detected by five serological methods including rose bengal plate test(RBT),standard agglutination test(SAT),competitive enzyme-linked immunosorbent assay(cELISA),indirect enzyme-linked immunosorbent assay(iELISA)and fluorescence polarization test(FPA),as well as loop-mediated isothermal amplification(LAMP)and realtime fluorescence quantitative PCR pathogenic detection methods,respectively,to analyze and compare the specificity,sensitivity,misdiagnosis rate,omission diagnostic rate,coincidence rate and Kappa value of those methods. The results showed that,compared with SAT,the sensitivity,specificity and coincidence rate were listed from high to low for serological methods were iELISA、FPA、cELISA and RBT,specifically,all their sensitivities,specificities and coincidence rates were above 85.00%、97.60%、97.50% and Kappa values were ≥1.00;compared with PCR,LAMP was with lower sensitivity(9.09%),stronger specificity(99.65%)and higher coincidence rate(98.49%)with PCR. In conclusion,the above serological methods were sensitive and specific,with high coincidence rate and consistency,while the molecular biological methods were with strong specificity,which could avoid misdiagnosis and omission misdiagnosis to some extent. Therefore,an applicable detection method should be selected to purify brucellosis in cattle in combination with the contamination and immunity of the cattle and different stages of purification,otherwise,some deviations would be produced by only one method. It was recommended,in order to determine any presence of Brucella infection,to use iELISA or FPA or RBT for preliminary screening,cELISA or SAT for diagnosis,and then molecular biological detection for vaginal swabs or milk samples from positive cattle. A reference was thus provided to select an appropriate method to purify and detect brucellosis in immunized cattle.

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吴小梅,李建玲,王涛,蔡扩军,朱冠林,杨镇,马小娟,高敏,陈利苹,钟旗,李爱巧.牛布鲁氏菌A19号疫苗免疫血清几种检测方法的比较[J].《中国动物检疫》编辑部,2022,39(5):110-115.

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