建立微量空斑减少方法测定人血清H7N9禽流感病毒中和抗体水平  

Establishment of micro-plaque reduction assay to detect human serum neutralization antibody level against avian influenza A(H7N9) virus

作　　者：薄洪[1] 董丽波[1] 张烨[1] 董婕[1] 邹淑梅[1] 高荣保[1] 王大燕[1] 舒跃龙[1] 

机构地区：[1]中国疾病预防控制中心病毒病预防控制所卫生部医学病毒和病毒病重点实验室,北京100026

出　　处：《疾病监测》2016年第2期126-129,共4页Disease Surveillance

基　　金：十二五重大专项课题(No.2014ZX10004002)~~

摘　　要：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 micro-plaque reduction assay to detect human serum level of neutralizing antibody against avian influenza A（ H7N9） virus. Methods M DCK cells were infected with H7N9 virus,plaques were measured to determine the titer of the virus. The optimal infectious dose of the H7N9 virus was calculated,human sera in serial dilutions were added to the virus at optimal infectious dose,the number of plaques reduced was measured,and the neutralizing antibody titer to H7N9 virus was determined. Results Plaque reduction method was used to detect anti H7N9 neutralizing antibody level,which had no cross reaction with seasonal influenza,avian influenza H5N1pdm09,H9N2 subtype virus antibodies.M icro-plaque reduction method showed consistent results with micro-neutralization method. Conclusion M icro-plaque reduction method had good sensitivity and specificity in detecting human serum level of neutralizing antibody against avian influenza A（ H7N9） virus.

关 键 词：微量空斑减少 禽流感病毒 H7N9 中和抗体 

分 类 号：R511.7[医药卫生—内科学]