CAJ | 학술논문

目的了解北京市门头沟区手足口病患儿感染病毒的型别,探索用荧光定量RT-PCR法对手足口病患者咽拭子标本中肠道病毒71型(EV71)、柯萨奇病毒A16(CoxA16)型病毒载量进行定量检测的可行性.方法采用实时荧光RT-PCR体外扩增法对81例手足口病患儿咽拭子标本提取的RNA进行检测.结果 28例手足口病患者体内CoxA16病毒载量＞103 copies· ml-1.实时荧光定量RT-PCR法构建的标准曲线显示,样本阈值环数(Ct)值与病毒拷贝数的对数(log10)之间的相关系数为0.9998,相关性良好.4例手足口病患者咽拭子标本中EV71型病毒载量＞103 copies· ml-1.实时荧光定量RT-PCR法标准曲线显示,Ct值与病毒拷贝数的对数之间的相关系数为0.9996,相关性较好.结论门头沟区手足口病病原谱以CoxA 16型为主,34.6％的手足口病患儿CoxA 16型病毒载量＞ 103 copies· ml-1,4.9％的患儿EV71型＞103 copies· ml-1.实时荧光定量RT-PCR法对咽拭子标本中CoxA16、EV71核酸定量检测比较简便快速、结果直观、稳定性强,为进一步探讨患者体内病毒载量与临床症状的关系打下了良好的基础.
목적 료해북경시문두구구수족구병환인감염병독적형별,탐색용형광정량RT-PCR법대수족구병환자인식자표본중장도병독71형(EV71)、가살기병독A16(CoxA16)형병독재량진행정량검측적가행성.방법 채용실시형광RT-PCR체외확증법대81례수족구병환인인식자표본제취적RNA진행검측.결과 28례수족구병환자체내CoxA16병독재량＞103 copies· ml-1.실시형광정량RT-PCR법구건적표준곡선현시,양본역치배수(Ct)치여병독고패수적대수(log10)지간적상관계수위0.9998,상관성량호.4례수족구병환자인식자표본중EV71형병독재량＞103 copies· ml-1.실시형광정량RT-PCR법표준곡선현시,Ct치여병독고패수적대수지간적상관계수위0.9996,상관성교호.결론 문두구구수족구병병원보이CoxA 16형위주,34.6％적수족구병환인CoxA 16형병독재량＞ 103 copies· ml-1,4.9％적환인EV71형＞103 copies· ml-1.실시형광정량RT-PCR법대인식자표본중CoxA16、EV71핵산정량검측비교간편쾌속、결과직관、은정성강,위진일보탐토환자체내병독재량여림상증상적관계타하료량호적기출.
Objective To identify the viral infection types of the children with hand-foot-mouth disease in Mentougou district of Beijing using real-time fluorescenee quantitative reverse transcription-polymerase chain reaction (RT-PCR) for detection of EV71 and CoxA16.Methods RNA was extracted from the throat swab of the children with hand-foot-mouth disease.The extracted RNA was tested by real-time fluorescence quantitative RT-PCR.Results The CoxA16 viral load of 28 samples exceeded 103 copies· ml-1.The standard curve of the real-time fluorescence quantitative RT-PCR showed good correlation coefficient (r =0.9998) between the cycle threshold values and logarithm of the viral copy number.The EV71 viral load of 4 samples exceeded 103 copies-ml-1.The standard curve of the real-time fluorescence quantitative RT-PCR showed good correlation coefficient (r =0.9996) between the cycle threshold values and logarithm of the viral copy number.Conclusions In this study,the main pathogenic detection was CoxA16 of hand-footmouth disease in Mentougou district of Beijing,34.6％ of the children with hand-foot-mouth disease were caused by CoxA16,4.9％ of the children with hand-foot-mouth disease were caused by EV71 in Mentougou district.Real-time fluorescence quantitative RT-PCR offers a rapid and simple method to detect EV71 or Cox A16 from specimens,and allow to analyze the association of the clinical symptoms and virus load.