猴源细小病毒血凝和血凝抑制试验的优化与应用

Optimization and application of hemagglutination and hemagglutination inhibition assays for detection of parvovirus in monkeys

目的优化猴源细小病毒的血凝和血凝抑制(HA/HI)试验条件,以优化的方法对160份2010年～2011年间采集于中国北京地区圈养猴群的血清样品进行猴源细小病毒抗体调查。方法 在不同的缓冲液、缓冲液pH值、猪红细胞浓度、兔血清浓度、阿氏液比例和温度下进行猴源细小病毒的HA/HI试验,选择合适的HA/HI条件。通过优化的HA/HI方法对160份猴血清进行猴源细小病毒抗体检测。结果pH 7.0、0.02 mol/L PBS、0.75%猪红细胞、0.5%兔血清和4℃可作为猴源细小病毒HA/HI试验合适的反应条件,猪血球以1∶1阿氏液抗凝,可保存5～7 d不影响实验结果。猴血清的猴源细小病毒抗体阳性率为58.1%。结论方法优化后稳定性增强、检测时间缩短、准确性提高。细小病毒在我国北京地区圈养猴群中感染比较普遍。

Objective To optimize the conditions of hemagglutination and hemagglutination inhibition（HA and HI） assays for detection of parvovirus in monkeys.Methods Hemagglutination and hemagglutination inhibition assays were carried out using different buffer solutions,buffer pH values,porcine erythrocyte concentrations,rabbit serum concentrations,proportion of Alsever＇s solution,and temperatures to detect monkey parvovirus titers,and to determine the optimal conditions of HA and HI asssays.A total of 160 sera samples were collected from several captive monkey centers in Beijing area during 2010 to 2011 and used to conduct the serological survey detecting the antibodies to simian parvovirus.Results 0.02 mol/L PBS at pH 7.0,0.75% porcine erythrocytes containing 0.5% rabbit serum and 4℃ temperature were confirmed to be optimal conditions.Porcine erythrocytes conserved in Alsever＇s solution at a ratio of 1：1 could be used for HA/HI assays within 7 days and was as sensitive as the fresh porcine erythrocytes.The positive rate of simian parvovirus-specific antibody was 58.1%.Conclusions Parvovirus infection is rather common in captive monkey populations in Beijing area in China.The improved hemagglutination inhibition assay is more accurate and stable,and less time comsuming than the currently used detection methods.