检测禽副粘病毒2型抗原的双抗体夹心ELISA方法的研究

Development of a Doule-antibody Sandwich ELISA to Detect Avian Paramyxovirus Type 2 Antigens

以禽副粘病毒 2型 (Yucaipa株 )群特异性单抗 7E5为基础建立了检测PMV_2抗原的双抗体夹心ELISA方法。该方法对Yucaipa病毒的最低检出量为 1.6 96 2 μg/ml,对PMV_2不同形式病毒及各分离株也都有很好的检出率。用Yucaipa病毒尿囊液人工感染 10日龄SPF鸡 ,于感染后不同时间采集心、肝、脾、肺、肾、法氏囊、气管等病科及泄殖腔拭子 ,经处理后用建立的方法进行检测 ,结果表明PMV_2病毒在感染鸡的法氏囊组织内大量存在 ,感染后 3天就可从法氏囊组织内检测到PMV_2病毒 ,粪便中仅个别有检出阳性者。

A double_antibody sandwich ELISA (DAS_ELISA)emplying a monoclonal antibody 7E 5 was developed for detection of PMV_2 antigen,the limit of viral antigens detected was 1.6962μg/ml.Using the DAS_ELISA detected standard virus and 3 Chinese isolates,the results indicated that the method had broad specificity.SPF chickens of 10 days old were artificially inoculated PMV_2(Yucaipa)by nasals and eyes,the chickens were killed at days of 1?2?3?5?7?9?11?13?15 after infection,and the tissues of liver and cloacal bursa were used for detection of PMV_2.The results confirmed that there were a lot of PMV_2 in cloacal bursa,and eloacal bursa was good tissue for the isolation of PMV_2.