流式细胞术检测人巨细胞病毒活动性感染检测方法的建立及检测效果评价  

Performance of Flow Cytometry for Detecting Active Infection of Human Cytomegalovirus and Evaluation of Its Efficacy

作　　者：王建[1] 曹伟锋 毛月燕[1] 吴力丰 庄国华[1] 钱春艳[1] 

机构地区：[1]杭州市余杭区第一人民医院,杭州311100 [2]浙江省药品化妆品审评中心,杭州310012

出　　处：《病毒学报》2017年第3期450-454,共5页Chinese Journal of Virology

基　　金：国家自然科学基金(项目号:81601831);题目:HCMV UL138在人巨细胞病毒潜伏感染中的作用及调控机制研究;杭州市医疗卫生科研项目(项目号:20150633B66);题目:HCMV潜伏感染与细胞自噬的相关性及IE1/Beclin 1通路的作用机制研究;杭州市余杭区医疗卫生重点科研项目(项目号:2015008);题目:HCMV潜伏感染与细胞自噬的相关性及IE1/Beclin 1通路的作用机制研究~~

摘　　要：探讨流式细胞术检测人巨细胞病毒活动性感染检测方法的可行性及效果评价。分离人外周血白细胞,以商品化的小鼠抗人巨细胞病毒pp65抗原单克隆抗体为一抗,FITC标记的羊抗小鼠IgG抗体为二抗,采用流式细胞术对外周血人巨细胞病毒pp65抗原进行检测。同时采用间接免疫荧光法对外周血人巨细胞病毒pp65抗原进行检测。采用配对χ2检验对两种方法的检测效果进行评价。临床送检的65份疑似为人巨细胞病毒感染病人外周血标本中,间接免疫荧光法检出阳性9份,流式细胞术检出阳性11份,两种方法阳性检出率差异无统计学意义(P>0.05)。采用流式细胞术可定量检测外周血人巨细胞病毒pp65抗原,与间接免疫荧光法检测结果无统计学差异,可在临床推广使用。To evaluate the feasibility and effectiveness of flow cytometry in detection of active infection of human cytomegalovirus （HCMV）, peripheral blood white blood cells were isolated, and human cytomegalovirus pp65 antigen was detected using flow cytometry（FCM） based on commercial mouse anti-human cytomegalovirus pp65 antigen monoclonal antibody as primary antibody and FITC-conjugated goat anti-mouse IgG antibody as secondary antibody. In addition, human cytomegalovirus pp65 antigen was detected in peripheral blood using indirect immunofluorescence assay（IFA）. The diagnostic efficacy of these two methods was compared using a paired chi-square test. Of 65 peripheral blood specimens of patients suspected of HCMV infections, IFA detected 9 positive specimens, whereas FCM detected 11 positive specimens. There was no significant difference in the detection rate between these two methods （P〉0.05）. FCM can be used to quantitatively detect human cytomegalovirus pp65 antigen in peripheral blood, and it has a high coincidence with IFA. FCM deserves popularization in the detection of HCMV active infection.

关 键 词：人巨细胞病毒(HCMV) PP65抗原 流式细胞术 间接免疫荧光法(IFA) 

分 类 号：R373.1[医药卫生—病原生物学]