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目的建立微量空斑减少的方法测定人血清中H7N9禽流感病毒中和抗体水平。方法确定H7N9禽流感病毒感染MDCK细胞后形成空斑数目,进行病毒定量,系列稀释待测抗体,加入已定量病毒,计算病毒被抗体中和后空斑减少数目,确定中和抗体的滴度。实验中以H7N9病例血清测定该方法的灵敏度,以其他亚型流感病毒抗体阳性人群血清评估微量空斑减少方法的特异性,同时将该方法与微量中和实验进行比较分析。结果微量空斑减少方法不仅能够检测H7N9禽流感病例中和抗体水平,而且该方法检测季节性流感病毒抗体、H5N1和H9N2禽流感病毒抗体与H7N9禽流感病毒抗体之间均无交叉反应,与微量中和实验检测结果相一致。结论建立的微量空斑减少方法具有较好的灵敏度和特异性,能够用于人血清H7N9禽流感病毒中和抗体的检测。
Objective To establish a method for the determination of neutralizing antibodies to H7N9 avian influenza virus in human serum by using the method of minimizing plaque reduction. The number of plaques was determined after infection with H7N9 avian influenza virus in MDCK cells. The virus was quantified and the antibody to be tested was serially diluted. The virus was added to quantify the number of viruses and the number of plaque reduction after antibody neutralization was determined. The titer of neutralizing antibody was determined. In the experiment, the sensitivity of this method was determined by the serum of H7N9 cases, and the specificity of the method for reducing plaque reduction was evaluated by the sera of other subtype influenza virus antibody positive people. Meanwhile, the method was compared with the micro-neutralization experiment. Results The micro-plaque reduction method not only detected neutralizing antibodies in H7N9 avian influenza cases, but also detected the antibody against seasonal influenza virus. There was no cross-reaction between H5N1 and H9N2 avian influenza virus antibodies and H7N9 avian influenza virus antibody, Neutralization test results are consistent. Conclusion The established method of reducing trace plaques has good sensitivity and specificity and can be used to detect the neutralizing antibodies of human serum H7N9 avian influenza virus.