PCR与ELISA和AGID间接检测牛白血病病毒的比较研究。

D. Martin, A. Arjona, I. Soto, N. Barquero, M. Viana, E. Gómez-Lucia
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引用次数: 22

摘要

诊断方法的选择取决于待分析畜群的特征。采用琼脂凝胶免疫扩散法(AGID)、酶联免疫吸附法(ELISA)和聚合酶链反应法(PCR)对2个不同流行性牛地方性白血病的牛群进行一致性研究。PCR是一种越来越常用的病毒学方法,用四组引物扩增不同的基因组区域(env, pol和tax),从外周血单核细胞(PBMCs)或乳汁白细胞中提取DNA。用引物扩增env或pol对pbmc中提取的DNA进行PCR,获得的阳性动物比例最高,其次是pbmc和与tax杂交的引物进行PCR,然后是血清ELISA,最后是AGID。与乳白细胞相比,PCR结果与pbmc更一致。
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Comparative study of PCR as a direct assay and ELISA and AGID as indirect assays for the detection of bovine leukaemia virus.
The choice of a diagnostic method depends on the characteristics of the herd to be analysed. Two herds with different prevalences of enzootic bovine leukaemia were chosen to study the concordance between agar gel immunodiffusion (AGID), enzyme-linked immunosorbent assay (ELISA) and polymerase chain reaction (PCR) methods. PCR, an increasingly used virological method, was performed with four sets of primers, amplifying different genomic regions (env, pol and tax), from DNA extracted either from peripheral blood monocytes (PBMCs) or milk leucocytes. The highest percentage of positive animals was obtained using PCR performed with DNA extracted from PBMCs using primers which amplified either env or pol, followed by PCR using PBMCs and primers which hybridized with tax, then ELISA using serum and finally AGID. The results of PCR were more consistent with PBMCs than when milk leucocytes were used.
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