单抗免疫过氧化物酶技术检测鸡传染性支气管炎病毒

A Monoclonal Antibody Immunoperoxidase Technique for Detection of Avian Infectious Bronchitis Virus

以抗鸡传染性支气管病毒（ＩＢＶ）核衣壳蛋白（Ｎ）的单抗株６ＤＨ８作为一抗，以辣根过氧化物酶标记的羊抗鼠ＩｇＧ作为二抗，建立了检测石蜡切片中ＩＢＶ抗原的单抗免疫过氧化物酶技术（Ｍｃ－ＩＰ），并对人工攻毒鸡及临床ＩＢＶ感染疑似鸡进行了检测。在ＩＢＶＭ４１株人工攻毒鸡，用该技术于１～１２ｄ从气管、２～７ｄ从肾脏可以检测到ＩＢＶ抗原，阳性染色集中于气管粘膜上皮细胞及肾小管上皮细胞胞浆；临床疑为ＩＢＶ感染的病鸡，以Ｍｃ－ＩＰ技术和单抗免疫荧光试验（Ｍｃ－ＩＦＡ）同时进行检测，结果阳性率分别为９０．３％及８３．９％。

An immunoperoxidase staining technique based on monoclonal antibody was developed and applied to detect avian infectious bronchitis virus (IBV) antigen in paraffin embedded tissue sections. The McAb used as primary antibody was specific to nucleocapsid protein of IBV and could react with a broad spectrum of homogeneous and heterogeneous strains. IBV antigen could be detected in chickens artificially inoculated with IBV strain M41 from tracheas on 1 12 days and kidneys on 2 7 days, and the positive staining was mainly observed in the epithelial cells of tracheal mucosas and uriniferous tubules. Thirty one chickens suspected to have infectious bronchitis were detected simultaneously by Mc IP staining and Mc IFA, positive rate of Mc IP staining (90.3%) being higher than that of Mc IFA (83.9%).