间接法Dot-ELISA检测鸡减蛋综合症抗体的建立及应用研究

Studies on Establishment and Application o the Detection of Antibodies to Egg Drop Syndrome-1976 by Indirect Dot-ELISA

应用ＳｅｐｈａｄｃｘＧ－２００层析法纯化鸡减蛋综合症病毒，利用ＮＣ膜作为载体，成功建立了检测ＥＤＳ－７６血请抗体的斑点酶联免疫吸附试验（Ｄｏｔ－ＥＬＩＳＡ）。抗原包被浓度为２μｇ／ｍｌ，被检血清浓度为１：２０，酶标兔抗鸡１ｇＧ浓度为１：２００．底物溶液最适ｐＨ值为７．２。对Ａ－Ｆ６个养禽场随机抽检血清１６０份，分别用Ｄｏｔ－ＥＬＩＳＡ、ＨＩ和ＡＧＰ检测，Ｄｏｔ－ＥＬＩＳＡ检出阳性率为５１．９％，ＨＩ检出阳性率为４６．９％，ＡＧＰ检出阳性率为３１．９％。对１４０日龄鸡人工感染试验，测定抗体消长规律。本方法不需特殊仪器。适用于基层兽医部门和养鸡场对ＥＤＳ－７６的血清学诊断和流行病学调查。

A indivect Dot - ELISA was developed for the detection ofantibodis against Egg Drop Syn- drome-1976(EDS-76). ED5-76 Virus (AV127)was purified by Sephadex G-200. lt was shown that coating concentrationwas 2 ug/ml. Optimal dilution of serum samples was 1 :20. Optimal dilutionof rab- bit anti-chicken antibodies conjugated by peoidase was 1: 200. 160serum samples from avian farm were detected by indirect Dot ELISA , Hland AGP ,the result indicateed that positive rates were 51. 9% , 46. 9%and 31. 9% ,respectiely, Titres of antibody in the serum of experimentalinfected chicken according to days were detected. It. was demonstratedthat the indirect Dot-ELISA was a specific ,sensitive ,rapid ,re- producibletest for serological survey of EDS - 76.