摘要
目的比较不同的脊髓灰质炎病毒(Poliovirus,PV,以下简称脊灰病毒)检测方法,为选择合理的优化检测方案提供参考。方法选取广西壮族自治区柳州市1200名2~3月龄健康婴儿,分为IPV和OPV不同序贯免疫共12组,每组100人,按照0、28、56d免疫程序接种疫苗,随机抽取各组入组时前10%的受试对象在第二剂疫苗接种前和第二、三剂疫苗接种后第7、14、28d采集粪便标本,利用实时荧光定量PCR和细胞培养法对粪便中可能存在的PV进行初筛对比,对细胞培养法分离出的阳性标本,分别用中和试验、单克隆抗体间接ELISA法和逆转录-PCR鉴定其型别,并比较3种方法分型结果的一致性。结果实时荧光定量PCR检测PV阳性率为76.4%,细胞培养法为59.7%,差异有统计学意义(χ^2=32.998,P<0.01)。中和试验、ELISA、RT-PCR 3种方法对细胞培养物中PV的检出率具有高度的一致性(Kappa>0.8)且具有较高的鉴别诊断准确性(ROC曲线面积A接近1),分型结果差异无统计学意义(P>0.05)。结论实时荧光定量PCR法对粪便中PV的检出率较细胞培养法高,但不能判断PV是否具有感染性。中和试验、ELISA、RT-PCR 3种方法对PV的分型一致性较高,可根据实验室和临床需求合理选择使用。
Objective To compare the different methods of detecting poliovirus in order to provide a reference for the selection of rational methods of detecting poliovirus. Methods A total of 1 200 healthy infants aged 2-3 months in Liuzhou,Guangxi Zhuang Autonomous region were randomly divided into 12 groups with different schedules for immunization with the inactivated polio vaccine(IPV)and the oral polio vaccine(OPV),and each group was vaccinated according to an immunization protocol of 0,28,and 56 days.Stool samples were collected before the second round of vaccination and on the 7 th,14 th,and 28 th day after the second and third rounds of vaccination in the first 10% of subjects randomly selected from each group.The fecal samples were screened using real-time fluorescence quantitative PCR and cell culture.The types of poliovirus in the supernatant of fecal samples isolated via cell culture were identified using a neutralization test,ELISA,or a reverse transcription polymerase chain reaction(RT-PCR).The consistency of the classification results of the three methods was compared. Results Poliovirus was detected with real-time quantitative PCR at a rate of 76.4%,and with cell culture at a rate of 59.7%;,the rate of detection differed significantly(χ^2=32.998,P<0.01).The rates of poliovirus detection via a cell culture involving the neutralization test,ELISA,or RT-PCR were highly consistent(Kappa>0.8)and had high diagnostic accuracy(the area under the ROC curve was close to 1).There were no significant differences in typing results(P>0.05). Conclusion The rate of poliovirus detection in original fecal samples according to real-time fluorescence quantitative PCR was higher than that with a cell culture,but PCR could not determine whether the virus is infective or not.A neutralization test,ELISA,and RT-PCR are highly consistent in terms of PV typing,and can be rationally selected and used according to testing and clinical needs.
作者
施红媛
傅宇婷
马汝飞
李琼芬
李国良
张杰
刘小畅
叶慧
王竞冬
石昊昱
杨晓蕾
杨净思
SHI Hong-yuan;FU Yu-ting;MA Ru-fei;LI Qiong-fen;LI Guo-liang;ZHANG Jie;LIU Xiao-chang;YE Hui;WANG Jing-dong;Shi Hao-yu;YANG Xiao-lei;YANG Jing-si(Institute of Med-ical Biology,Chinese Academy of Medical Sciences,Peaking Union Medical College,Kuming,Yunnan,China 650118;Yunnan Center for Disease Control and Prevention,Kunming,Yunnan,China 650032;Tianjin Center for Disease Control and Prevention,Tianjin,China 300011;Hangzhou Women's Hospital,Hangzhou)
出处
《中国病原生物学杂志》
CSCD
北大核心
2020年第8期947-952,共6页
Journal of Pathogen Biology
基金
国家"重大新药创制"科技重大专项(No.2015ZX09101031,2016ZX09106003-007)
云南省重点新产品开发专项(No.2016BC002)
工信部"2019年儿童用联合疫苗、肺炎结合疫苗等新型疫苗项目"(No.0714-EMTC-02-00908)。
关键词
脊髓灰质炎
病毒检测
方法比较
poliomyelitis
virus detection
comparison of methods
