免疫酶技术检测鸡传染性支气管炎病毒的研究

DETECTION OF AVIAN NEPHROPATHOGENIC INFECTIOUS BRONCHITIS VIRUS BY IMMUNOHISTOCHEMICAL STAINING

以抗鸡肾型传染性支气管炎病毒（肾型ＩＢＶ）Ｔ株的鼠源高免血清为一抗，辣根过氧化物酶（ＨＲＰ）标记的羊抗鼠ＩｇＧ为二抗，建立了检测石蜡切片中肾型ＩＢＶ抗原的免疫酶组化技术，经过特异性试验和阻断性试验，表明该方法具有高度的敏感性和特异性．应用该法对人工感染肾型ＩＢＶＲＳ株的鸡气管和肾脏石蜡切片中的病毒抗原进行了检测，结果气管从感染后０．５～１１ｄ、肾脏从感染后１～２０ｄ均检测到病毒抗原。

An immunohistochemical technique based on hyperimmune serum was developed and applied to detect avian nephropathogenic infectious bronchitis virus antigens in paraffin embeded tissue sections. Optimal dilution titers of the hyperimmune serum and HRP conjugated goat anti mouse antibodies were 1∶100 and 1∶400, respectively. The results of the specificity and blocking tests showed that this technique had good sensitivity and high specificity. With this prcedure, nephropathogenic infectious bronchitis virus antigens could be detected from 12 hours to 11 days in chachea and from 24 hours to 20 days in kidney post infection. Positive staining was mainly observed in the epithelia of the tracheal mucosa, renal tubules and ureter.