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不同分子检测策略诊断手足口病住院患儿肠道病毒的一致性

张天琛,  杨俊梅,  李昱,  杨建丽,  周永红,  康迪,  邱琪,  李鹏,  梁露,  申远方,  崔鹏,  崔亚杰,  郭淳,  宋春兰,  曾梦遥,  王芳,  龙璐,  成怡冰,  余宏杰

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文章导航 > 中华疾病控制杂志  > 2020 >  24(10): 1202-1208
张天琛, 杨俊梅, 李昱, 杨建丽, 周永红, 康迪, 邱琪, 李鹏, 梁露, 申远方, 崔鹏, 崔亚杰, 郭淳, 宋春兰, 曾梦遥, 王芳, 龙璐, 成怡冰, 余宏杰. 不同分子检测策略诊断手足口病住院患儿肠道病毒的一致性[J]. 中华疾病控制杂志, 2020, 24(10): 1202-1208. doi: 10.16462/j.cnki.zhjbkz.2020.10.018
引用本文: 张天琛, 杨俊梅, 李昱, 杨建丽, 周永红, 康迪, 邱琪, 李鹏, 梁露, 申远方, 崔鹏, 崔亚杰, 郭淳, 宋春兰, 曾梦遥, 王芳, 龙璐, 成怡冰, 余宏杰. 不同分子检测策略诊断手足口病住院患儿肠道病毒的一致性[J]. 中华疾病控制杂志, 2020, 24(10): 1202-1208. doi: 10.16462/j.cnki.zhjbkz.2020.10.018
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ZHANG Tian-chen, YANG Jun-mei, LI Yu, YANG Jian-li, ZHOU Yong-hong, KANG Di, QIU Qi, LI Peng, LIANG Lu, SHEN Yuan-fang, CUI Peng, CUI Ya-jie, GUO Chun, SONG Chun-lan, ZENG Meng-yao, WANG Fang, LONG Lu, CHENG Yi-bing, YU Hong-jie. Estimation of the agreement between different molecular detection strategies for diagnosing Enterovirus in hospitalized children with hand, foot and mouth disease[J]. CHINESE JOURNAL OF DISEASE CONTROL & PREVENTION, 2020, 24(10): 1202-1208. doi: 10.16462/j.cnki.zhjbkz.2020.10.018
Citation: ZHANG Tian-chen, YANG Jun-mei, LI Yu, YANG Jian-li, ZHOU Yong-hong, KANG Di, QIU Qi, LI Peng, LIANG Lu, SHEN Yuan-fang, CUI Peng, CUI Ya-jie, GUO Chun, SONG Chun-lan, ZENG Meng-yao, WANG Fang, LONG Lu, CHENG Yi-bing, YU Hong-jie. Estimation of the agreement between different molecular detection strategies for diagnosing Enterovirus in hospitalized children with hand, foot and mouth disease[J]. CHINESE JOURNAL OF DISEASE CONTROL & PREVENTION, 2020, 24(10): 1202-1208. doi: 10.16462/j.cnki.zhjbkz.2020.10.018
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不同分子检测策略诊断手足口病住院患儿肠道病毒的一致性

doi: 10.16462/j.cnki.zhjbkz.2020.10.018
  • 1.

    102206 北京, 中国疾病预防控制中心传染病预防控制处/传染病监测预警国家重点实验室

  • 2.

    330029 南昌, 江西省疾病预防控制中心应急办

  • 3.

    450003 郑州, 郑州大学附属儿童医院/河南省儿童医院临床检验科

  • 4.

    450003 郑州, 郑州大学附属儿童医院/河南省儿童医院急危重症医学重点实验室

  • 5.

    200032 上海, 复旦大学公共卫生学院/公共卫生安全教育部重点实验室

  • 6.

    450003 郑州, 郑州大学附属儿童医院/河南省儿童医院感染性疾病科

  • 7.

    610041 成都, 四川大学华西公共卫生学院

  • 8.

    430030 武汉, 华中科技大学同济医学院公共卫生学院流行病与卫生统计系

张天琛与杨俊梅为共同第一作者.
基金项目: 

国家“艾滋病和病毒性肝炎等重大传染病防治”重大专项 2017ZXl0103009-005

国家杰出青年科学基金“传染病流行病学” 81525023

艾滋病和病毒性肝炎等重大传染病防治”重大专项 2018ZXl0713001

艾滋病和病毒性肝炎等重大传染病防治”重大专项 2018ZXl0201001-010

艾滋病和病毒性肝炎等重大传染病防治”重大专项 2018ZXl0713001.007

详细信息
    通讯作者:

    成怡冰, E-mail:13703829317@163.com

    余宏杰, E-mail:yhj@fudan.edu.cn

  • 中图分类号: R512.57;R373.25

Estimation of the agreement between different molecular detection strategies for diagnosing Enterovirus in hospitalized children with hand, foot and mouth disease

  • 1.

    Division of Infectious Diseases, Chinese Center for Disease Control and Prevention/Key Laboratory of Surveillance and Early Warning on Infectious Disease, Beijing 102206, China

  • 2.

    Department of Emergency Management, Jiangxi Center for Disease Control and Prevention, Nanchang 330029

  • 3.

    Department of Clinical Laboratory, Children's Hospital Affiliated to Zhengzhou University/Children's Hospital of Henan Province, Zhengzhou 450003, China (Yang JM, Yang JL, Kang D)

  • 4.

    Key Laboratory of Critical Care Medicine, Children's Hospital Affiliated to Zhengzhou University/Children's Hospital of Henan Province, Zhengzhou 450003, China

  • 5.

    School of Public Health, Fudan University/Key Laboratory of Public Health and Safety Education Ministry, Shanghai 200032, China

  • 6.

    Department of Infectious Diseases, Children's Hospital affiliated to Zhengzhou University/Children's Hospital of Henan Province, Zhengzhou 450003, China

  • 7.

    West China School of Public Health, Sichuan University, Chengdu 610041, China

  • 8.

    Department of Epidemiology and Biostatistics, School of Public Health, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China

ZHANG Tian-chen and YANG Jun-mei contributed equally to this article
Funds: 

National Science and Technology Major Project of China 2017ZXl0103009-005

The National Science Fund for Distinguished Young Scholars "Infectious Disease Epidemiology" 81525023

National Science and Technology Major Project of China 2018ZXl0713001

National Science and Technology Major Project of China 2018ZXl0201001-010

National Science and Technology Major Project of China 2018ZXl0713001.007

More Information

    摘要
  • 摘要:   目的  本研究旨在了解单纯实时荧光定量逆转录聚合酶链式扩增技术（fluorescence-based quantitative real-time reverse transcription-polymerase chain reaction，qRT-PCR）及qRT-PCR联合巢式聚合酶链式反应（nested polymerase chain reaction，nested PCR）测序两种检测策略在检测手足口病（hand，foot and mouth disease，HFMD）住院患儿肠道病毒的一致性及其相关因素。  方法  2017年2月―2018年2月在河南省建立以医院为基础的HFMD住院患儿前瞻性队列，收集研究对象相关信息并采集生物标本，分别开展单纯qRT-PCR及qRT-PCR联合nested PCR测序的肠道病毒核酸检测。  结果  两种检测策略总体的一致性较好（kappa>0.60）。标本类型、肠道病毒A组71血清型（enterovirus-A71，EV-A71）疫苗接种史、发病到采样时间、疾病严重程度以及年龄与肠道病毒的检出收益相关。qRT-PCR联合nested PCR测序的样本采样时间≥4 d、不同检测策略的样本采样时间差>2 d会降低两种检测策略EV-A71矫正机遇后的一致性，而重症与EV-A71矫正机遇后的高一致性有关。标本类型（P=0.033）及疾病严重程度（P=0.020）是两种检测策略柯萨奇病毒A组16（coxsackievirus-A16，CV-A16）矫正机遇后一致性的独立相关因素。  结论  qRT-PCR及qRT-PCR联合nested PCR测序的两种检测策略的一致性较好，患儿个体情况及标本采集时间会影响分子检测的检出收益及一致性。
    Abstract:   Objective  The aim of this study was to understand the diagnostic yield of fluorescence-based quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR) only and qRT-PCR combined with several nested polymerase chain reaction (nested PCR) sequencing for detecting enterovirus in hospitalized children with hand, foot and mouth disease (HFMD), furthermore to analyze the agreement and its associated factors of the two testing strategies.  Methods  A hospital-based prospective HFMD cohort was established in Henan from February 2017 to February 2018. The relevant information and biological specimens of the participants were collected. According to the detection strategies, qRT-PCR and qRT-PCR combined with several nested PCR sequencing were performed both for enterovirus diagnosis independently.  Results  The agreement between the two testing strategies in the detection for enterovirus-A71 (EV-A71), coxsackievirus-A16 (CV-A16) and other enterovirus (EVs) were good (kappa>0.60). Sampling types and with EV-A71 vaccination, the time of sample collection after onset, age and disease severity were related to detection yield. The sampling days after onset of specimens for qRT-PCR combined with nested PCR sequencing ≥ 4 d and the sampling time interval of different detection strategies more than 2 d would reduce the chance-corrected agreement between the two testing strategies in EV-A71 detection. Severe disease was associated with higher chance-corrected agreement in EV-A71 detection. Specimen type (P=0.033) and disease severity (P=0.020) were independent factors related to the chance-corrected agreement of CV-A16 detection.  Conclusions  The agreement between the two detection strategies of qRT-PCR and qRT-PCR combined with nested PCR sequencing is relative well. The individual condition of the child and the time of specimen collection would affect the detection yields and consistency of molecular detection.
    • HTML全文

  • 图  1  两种检测策略肠道病毒检出率的比较

    Figure  1.  Comparison of enteroviruses detection yields between two testing strategies

    下载: 全尺寸图片 幻灯片

    图  2  两种检测策略不同标本及不同采样时间的肠道病毒检出率比较

    图标A、B、C分别代表总肠道病毒的检出率、EV－A71的检出率及CV－A16的检出率图标1、2分别代表qRT－PCR联合nested PCR测序策略和商品化试剂盒qRT－PCR检测策略

    Figure  2.  Enteroviruses detection yields of two testing strategies in different sample types and sampling days after onset

    下载: 全尺寸图片 幻灯片

    图  3  不同标本及不同诊断结局的两种检测策略一致性分析

    注:Bangdiwala图可以直观显示一致性的关键信息ꎮ一致程度由位于对角线的黄色矩形内的蓝色框面积表示.黄色矩形表示最大可能的一致性, 其中黄色框中的蓝色框所占面积越大表示实际观察到的一致性越高.图标的A、B、C、D分别表示:(A)检测结果分为5类(B)按EV －A71阴、阳性分类(C)按CV－A16阴、阳性分类(D)按其他EVs阴、阳性分类.图标的1, 2, 3分别表示按照开展qRT－PCR联合nested PCR测序检测的样本类型分为:(1)全部样本; (2)咽拭子样本; (3)直肠样本.

    Figure  3.  Agreement analysis of two detection strategies for different specimens and different diagnosis outcomes

    下载: 全尺寸图片 幻灯片

    表  1  影响两种检测策略肠道病毒检出收益的多因素分析结果(基于Logistic回归分析模型)

    Table  1.   Associated factors of enterovirus detection yield for two testing strategies (based on Logistic regression analysis model)

    检测策略 检测结果(阳性vs.阴性) ORadj(95% CI)值
    标本类型(咽拭子vs.直肠标本) 发病至采样时间
    (≥4 d vs.＜4 d)    		 疾病严重程度(重症vs.轻症) 年龄
    (＞2岁vs.≤2岁)    		 EV-A71疫苗接种史(是vs.无)
    qRT-PCR+nested EV-A71 0.25(0.14，0.44)a 1.79(1.26，2.57)a 4.38(3.11，6.16)a 2.19(1.56，3.09)a 0.43(0.20，0.80)b
    PCR测序检测 CV-A16 0.38(0.22，0.68)a 0.69(0.51，0.92)b 0.30(0.18，0.48)a 1.17(0.86，1.59) 0.88(0.59，1.30)
    其他肠道病毒 1.82(1.11，2.99)a 0.61(0.49，0.76)a 0.70(0.54，0.90)a 0.62(0.49，0.78)a 1.53(1.11，2.12)a
    商品化试剂盒的 EV-A71 - 3.15(2.15，4.75)a 5.79(4.22，7.98)a 1.83(1.32，2.53)a 0.16(0.06，0.37)a
    qRT-PCR检测 CV-A16 - 0.74(0.58，0.95)b 0.42(0.29，0.60)a 1.46(1.12，1.89)a 0.90(0.63，1.26)
    其他肠道病毒 - 0.74(0.59，0.93)b 0.48(0.37，0.63)a 0.54(0.43，0.68)a 1.74(1.27，2.42)a
    注：a表示P＜0.01，b表示P＜0.05。
    下载: 导出CSV

    表  2  影响两种检测策略修正机遇后一致性的多因素分析结果(基于两阶段的Logistic回归分析模型)

    Table  2.   Associated factors of chance-corrected agreement between two testing strategies(based on two-stages Logistic regression analysis model)

    病原检测一致性(一致vs.不一致) ORadj(95% CI)值
    检测方法A的样本类型(咽拭子vs.直肠标本) 检测方法A的发病到采样时间(≥4 d vs.＜4 d) 检测方法B的发病到采样时间(≥4 d vs.＜4 d) 两种检测方法采样时间差(0~2 d vs.＜0 d) 两种检测方法采样时间差(＞2 d vs.＜0 d) 疾病严重程度(重症vs.轻症) 年龄(＞2岁vs. ≤2岁) EV-A71疫苗接种史(是vs.无)
    EV-A71 1.61(0.85，2.93) 0.45(0.26，0.76)a 1.15(0.57，2.22) 0.69(0.44，1.08) 0.41(0.20，0.86)b 1.67(1.11，2.51)b 1.17(0.79, 1.75) 0.99(0.43, 2.87)
    CV-A16 0.33(0.10，0.81)b 0.84(0.55，1.28) 1.21(0.76，1.92) 0.83(0.56，1.21) 1.70(0.68，5.18) 0.59(0.38，0.94)b 0.80(0.58, 1.13) 0.84(0.55, 1.33)
    其他肠道病毒 0.80(0.40，1.47) 0.79(0.56，1.12) 1.14(0.76，1.71) 1.02(0.74，1.41) 0.58(0.33，1.05) 0.83(0.61，1.15) 1.03(0.77, 1.39) 0.76(0.52, 1.14)
    注：检测策略A：qRT-PCR联合nested PCR测序检测。检测策略B：商品化试剂盒的qRT-PCR检测。两种检测策略采样时间差：检测策略B的采样日期减去检测策略A的采样日期。a表示P＜0.01，b表示P＜0.05。
    下载: 导出CSV
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  • 修回日期:  2020-06-27
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