检验医学 ›› 2012, Vol. 27 ›› Issue (5): 416-418.

• 分子生物学检验论著 • 上一篇    下一篇

实时荧光聚合酶链反应检测患儿手足口病病原体

张群,杨蕾,曹科峰   

  1. 太和县中医院检验科,安徽 太和 236000
  • 收稿日期:2012-04-23 出版日期:2012-05-30 发布日期:2012-05-10
  • 作者简介:张 群,男,1974年生,主管技师,主要从事生化检验工作。

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

  1. Department of Clinical Laboratory, Taihe Traditional Chinese Medicine Hospital, Anhui Taihe 236000, China
  • Received:2012-04-23 Online:2012-05-30 Published:2012-05-10

摘要: 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。

关键词: 实时荧光聚合酶链反应, 手足口病, 肠道病毒

Abstract: 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 (P<0.05). 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.

Key words: Real-time fluorescence polymerase chain reaction, Hand-foot-mouth disease, Enterovirus