鹅细小病毒单抗双夹心ELISA检测方法的建立

Development of double sandwich ELISA for detecting goose parvovirus based on monoclonal antibodies

为建立鹅细小病毒(GPV)检测方法,本研究以免抗GPV IgG为捕获抗体,抗GPV单克隆抗体为检测抗体,通过方阵试验确定了兔抗GPV IgG最适包被浓度为16 mg/L,抗GPV单克隆抗体最佳工作浓度为10.8 mg/L,酶标二抗最佳稀释倍数为1:4 000,建立了检测GPV单抗双夹心ELISA方法,对GPV检测灵敏度达0.312 mg/L。用该法对延边某养鹅场疑似发生小鹅瘟病的252只病死鹅进行检测,结果阳性率为75.79%,与病毒中和试验方法的检测结果符合率达91.11%。本研究建立的单抗双夹心ELISA方法为GPV的检测和区域流行病学调查提供了一种简便快速的血清学诊断方法。

In order to establish the method for detecting GPV,research rabbit anti-GPV IgG was used to be capture antibody and anti-GPV monoclonal antibody was used to be detection of antibody.Based on chess-board experiment, the optimal coating concentration of rabbit anti-GPV IgG is 16 mg/L,the optimal working concentration of anti-GPV monoclonal antibody is 10.8 mg/L,The optimization of dilution for enzyme-labeled second antibody is 1：4 000,and double sandwich ELISA whose sensitivity of detecting GPV is 0.312 mg/L.was established.Dead gooses in a goose ranch in Yanbian were detected by this method.The positive rate is 75.79%.The concordant rate with virus neutralization test is 91.11%.Double sandwich ELISA established by this research provides a convenient and fast sero-diagnosis for detecting GPV and epidemiologic survey in district.