对鸡群A／B亚群禽白血病病毒抗体ELISA检测阳性率的可靠性评估

Reliability Evaluation of ELISA for Antibodies to ALV-A/B in Chicken Serum Samples

为了评估某些批次的禽白血病病毒(ALV)抗体商品化ELISA检测试剂盒在检测鸡群中A/B亚群禽白血病病毒(ALV-A/B)抗体阳性率的可靠性,使用同一批次试剂盒,对往年ALV-A/B抗体检测阳性率达标(<1%)的A类鸡场及往年ALV-A/B抗体检测阳性率不达标(>1%)的B类鸡场所送检样品进行初检,在A类鸡场初检阳性样品中挑选S/P值最高的8份阳性样品,在B类鸡场初检阳性样品中挑选S/P值最高的40份样品,再使用相同批次的试剂盒将每个初检阳性样品做3个孔的重复检测,并以上述48份样品为一抗同时做间接免疫荧光试验(IFA),系统比较了IFA和ELISA 2种方法检测结果的一致性。结果发现,A类鸡场初检为阳性的8份样品经复检和IFA检测均变为阴性;B类鸡场4份样品(4/40)ELISA复检和IFA结果均变为阴性,36份样品(36/40)ELISA复检仍为阳性,但其中20份呈IFA阳性,16份呈IFA阴性,结果表明某些批次的试剂盒在特异性和稳定性方面存在一定问题。提示,相关企业在进行大批量样本检测时,ELISA阳性样品需结合IFA进行结果判定,以提高检测的准确性。

To evaluate the reliability of the positive rate of ALV-A/B specific antibodies detection in chicken serum and illustrate the stability and specificity problems existing in the commercialized Avian Leukosis Virus Subgroup A/B Antibody Test Kit, two batches of serum samples submitted by the company A which had no positive samples detected previously and the company B which had positive samples detected yet were tested using the same lot of ELISA kit by the professional operators. Then, among the positive samples detected for the first time, eight samples from company A and forty samples from company B whose S/P ratio all ranked higher were re-detected triply, and IFA were tested using the forty-eight samples above as first antibodies. The consistence of the results between ELISA and IFA assay was compared. The results demonstrated that, eight samples from company A turned into negative in the re-detection by ELISA kit, and IFA results were negative as well, which indicated stability problems existing in some batches of commercialized kit. Four samples （4/40） form company B turned into negative in the re-detection by ELISA kit, so as IFA results; thirty-six samples （36/40） form company B were positive in the re-de tection by ELISA kit, however, twenty of them were IFA positive and sixteen of them were IFA negative, the complicated results indicated non-specificity detection were existed in some batches of commercialized kit except for the stability problem. Therefore, to achieve higher veracity, samples should be determined by both of ELISA and IFA in the massive detection.