手足口病病原体实时荧光RT—PCR反应体系的建立与临床应用  

Development and clinical application of fluorescent real-time RT-PCR to hand, foot and mouth disease

作　　者：张永乐[1] 潘克女[3] 徐岱[1] 梁伟峰[2] 娄国强[2] 

机构地区：[1]杭州市第六人民医院临床实验室,310014 [2]杭州市第六人民医院中西医结合科,310014 [3]杭州市第一人民医院检验科

出　　处：《中华微生物学和免疫学杂志》2009年第3期276-278,共3页Chinese Journal of Microbiology and Immunology

摘　　要：目的建立一种快速、准确、特异性高的方法检测手足口病病原体。方法根据引起手足口病常见致病原肠道病毒71（EV71）、柯萨奇病毒A16（CA16）及肠道病毒（EV），设计相应的引物、探针对35例临床诊断的手足口病患儿及20例正常健康婴儿的粪便进行EV、EV71、CA16三种病原体实时荧光RT-PCR检测，同时对55分标本进行EV71病毒培养分离。结果35例临床诊断手足口病患儿粪便中EV全部阳性，EV71阳性25例，CA16阳性8例，其中3例为EV71、CA16同时阳性，与临床诊断符合率为85．71％，20例健康体检婴儿粪便中EV病毒5例阳性，EV71、CA16均阴性。结论荧光RT-PCR对手足口病病原体检测准确性高、特异性强，具有快速、廉价等特点，适合于手足口病的早期诊断。Objective To develop a rapid, accurate, specific method to detect causative agent of hand, foot and mouth disease （ HFMD ）. Methods Specific primers and probe were designed based on highly conserved VP1 region of enterovirus 71, coxsackie virus A16 and enterovinls. The sensitivity and specificity of the real-time RT-PCR was evaluated with 35 stool samples collected from pediatric patients with suspected HFMD and 20 clinical samples from health pediatric patients. Results Out of 35 clinical samples from suspected HFMD, 35 samples were identified as positive for enterovirus, 25 clinical samples were identified as positive for enterovirus 71, 8 clinical samples were identified as positive for coxsackie virus A16, among which 3 clinical samples were identified as positive for enterovirus 71 and coxsackie virus A16. The clinical diagnostic accordance rate is 85.71%. Out of 20 clinical samples from normal pediatric patients, 5 clinical samples were identified as positive for enterovirus, 20 clinical samples were negative for enterovirus 71 and coxsackie virus A16. Conclusion Our results indicate real-time RT-PCR offers a rapid, sensitive, specific and cheap method to detect pathogen of HFMD from clinical specimens.

关 键 词：手足口病 肠道病毒 柯萨奇病毒A16 实时荧光RT—PCR 

分 类 号：R725.1[医药卫生—儿科] R450[医药卫生—临床医学]