酶联免疫吸附试验与实时荧光聚合酶链反应对A组轮状病毒的检测效果比较  

Comparison of enzyme-linked immunosorbent assay and real-time fluorescent polymerase chain reaction in the detection of rotavirus group A

作　　者：甄博珺[1] 田祎[2] 邹林[1] 高翔[1] 张萍[1] 张靖[1] 张扬[1] 郭晓晨 高志勇[2] ZHEN Bo-jun;TIAN Yi;ZOU Lin;GAO Xiang;ZHANG Ping;ZHANG Jing;ZHANG Yang;GUO Xiao-chen;GAO Zhi-yong(Laboratory of Tongzhou Center for Disease Control and Prevention,Beijing 101100,China;不详)

机构地区：[1]北京市通州区疾病预防控制中心检验科,北京101100 [2]北京市疾病预防控制中心传染病地方病控制所,北京100013

出　　处：《中国病毒病杂志》2023年第1期63-66,共4页Chinese Journal of Viral Diseases

基　　金：首都卫生发展科研专项项目(首发2020-2-1011)。

摘　　要：目的比较酶联免疫吸附试验(enzyme linked immunosorbent assay,ELISA)与实时荧光聚合酶链反应(real-time fluorescent polymerase chain reaction,实时荧光PCR)对A组轮状病毒的检测效果,为后续试验提供参考。方法收集2015—2020年北京市某医院肠道门诊5岁以下腹泻儿童粪便样本,使用ELISA和实时荧光PCR两种方法检测A组轮状病毒,采用GP基因分型方法检测ELISA和实时荧光PCR检测阳性的样本。使用SPSS 20.0软件进行Kappa和Spearman统计分析。结果2015—2020年共采集1412份粪便样本,其中155份ELISA检测阳性,166份实时荧光PCR检测阳性。对实时荧光PCR和ELISA两种方法的检测结果进行Kappa检验,Kappa=0.954>0.75,说明这两种诊断方法检测一致性良好。ELISA法的吸光度A值呈偏态分布,实时荧光PCR法的Ct值类似正态分布,Spearman相关系数r=0.088,P=0.278,Ct值和吸光度A值无相关性。有13份标本两种方法检测结果不一致,仅有1份为ELSIA阳性、实时荧光PCR阴性,12份均为ELSIA阴性、实时荧光PCR阳性,其中,10份有GP分型结果,4份为G2[P4],4份为G9[P8],1份为混合感染G2/9[P4],1份为未分型。结论两种诊断方法定性诊断的一致性良好,但ELISA吸光度A值和实时荧光PCR Ct值无相关性,不能通过任何一种方法结果数值来推断另一种方法的结果数值。基因分型结果提示ELISA阴性、实时荧光PCR阳性的样本可能与G2[P4]型别的毒株与检测抗体的匹配度不是很高有关。Objective To compare the efficacy of enzyme-linked immunosorbent assay(ELISA)and real-time fluo⁃rescent polymerase chain reaction(qPCR)in the detection of rotavirus A,and to provide a reference for subsequent experiments.Methods From 2015 to 2020,stool samples of children with diarrhea under 5 years of age in the gastrointestinal clinic of a hospital in Beijing were collected,and rotavirus A was detected parallelly by ELISA and qPCR,GP genotyping was further performed on positive samples.Kappa and Spearman in SPSS 20.0 software were used for data statistical analysis.Results A total of 1412 stool samples were collected,of which 155 were ELISA positive and 166 were qPCR positive.Kappa test showed a high consistency of the two methods(Kappa=0.954>0.75).The absorbance A value of ELISA showed a skewed distribution,while the Ct value of qPCR exhibited a normal distribution,Spearman test indicated an uncorrelated relationship between Ct value and absorbance A value(r=0.088,P=0.278).There were 13 specimens that had different results,one sample was positive for ELISA while negative for qPCR,12 were positive for qPCR while negative for ELISA,among which 10 samples had GP typing results,showing 4 were G2[P4],4 were G9[P8],one was mixed infection of G2/9[P4],and one was undetermined.Conclusions Although the results of ELISA and qPCR in the detection of rotavirus A are highly consistent,there is no correlation between ELISA A value and qPCR Ct value.Therefore,the result of one method may not be used to predict the result of another.

关 键 词：A组轮状病毒 ELISA 实时荧光PCR 一致性检验 相关性检验 GP基因分型 

分 类 号：R373.9[医药卫生—病原生物学]