实时荧光聚合酶链反应检测患儿手足口病病原体

目的采用实时荧光定量聚合酶链反应(FQ-PCR)检测患儿手足口病病原体,为防治手足口病提供依据。方法收集手足口病疑似患儿189例(按年龄分为<3岁组113例、3～5岁组76例)及39例手足口病密切接触儿童的咽拭子、疱疹液,用FQ-PCR检测标本中的肠道病毒通用型(EV)、肠道病毒71型(EV71)及柯萨奇病毒A组16型(CoxA16),并与酶联免疫吸附试验(ELISA)结果进行比对。结果 189例手足口病患儿疑似组FQ-PCR检测EV阳性141例,阳性率为74.6%,其中<3岁阳性91例、3～5岁阳性50例,3～5岁组EV检出率明显低于<3岁组(P<0.05);EV71阳性135例,CoxA16阳性65例;咽拭子标本阳性率为74.5%(120/161),疮疹液标本阳性率为82.1%(23/28);ELISA检测EV阳性76例(40.2%);39例手足口病密切接触者FQ-PCR检测EV阳性率为33.3%(13/39),ELISA检测阳性率为17.9%(7/39)。结论 FQ-PCR是一种特异性强、灵敏度高、检测较快速、效率高的方法,对检测手足口病病原体及进行相关流行病调研有重要价值。

Analysis of pathogen causing hand-foot-mouth disease by real-time fluorescence PCR

Objective To detect the pathogens by real-time fluorescence quantitation polymerase chain reaction（FQ-PCR） and provide the reference for the prevention of hand-foot-mouth disease.Methods Throat swab and vesicle fluid were collected from 189 suspected children with hand-foot-mouth disease and 39 children with close contact to hand-foot-mouth disease patients in Anhui county and its neighborhood area.Enterovirus（EV）,enterovirus 71（EV71） and coxsakievirus A16（CoxA16） were detected by FQ-PCR,and the results were compared with the results of enzyme-linked immunosorbent assay（ELISA）.Results By FQ-PCR,141 children among the 189 suspected children with hand-foot-mouth disease were detected EV（positive rate： 74.6%）,including 91 children between 0 and 3 years old and 50 children between 3 and 5 years old.EV detection rate of 3-5 years old group was obviously lower than that of 0-3 years old group（P0.05）.There were 135 EV71 positive samples and 65 CoxA16 positive samples.The positive rate of throat swab samples was 74.5%（120/161）,and that of vesicle fluid samples was 82.1%（23/28）.There were 76（40.2%） EV positive samples by ELISA.The positive rate of EV in 39 children with close contact to hand-foot-mouth disease patients by FQ-PCR was 33.3%（13/39）.The positive rate by ELISA was 17.9%（7/39）.Conclusions FQ-PCR has high specificity,sensitivity,efficiency and speed,which contributes to its important value as detection method of pathogen causing hand-foot-mouth disease and epidemiological study.