多重巢式PCR应用于1起聚集性发热疫情的病原诊断  

Application of multiplex nested PCR technology for identification and genetyping for adenovirus in a fever outbreak in Changping district

作　　者：吴杨[1] 高进玺[1] 庄鹏[1] 李东迅[1] 刘重程[1] 舒高林 牛桓彩[1] WU Yang;GAO Jin-xi;ZHUANG Peng;LI Dong-xun;LIU Zhong-cheng;SHU Gao-lin;NIU Huan-cai(Changping District Center for Disease Control and Prevention,Beijing 102200,China)

机构地区：[1]北京市昌平区疾病预防控制中心,北京102200

出　　处：《江苏预防医学》2018年第2期124-127,共4页Jiangsu Journal of Preventive Medicine

摘　　要：目的增强实验室对突发发热疫情的快速诊断能力,完善、优化聚集性发热的病原检测方法。方法采用多重巢式PCR扩增法,对1起聚集性发热疫情样本进行病原筛查,同时对扩增产物测序,进行BLAST比对,进行血清型分型,比较不同引物的扩增效果。结果采集7份病例咽拭子标本,以呼吸道病毒核酸多重联检试剂盒进行14种病毒筛查,其中4份为腺病毒阳性,阳性率为57.1%。采用3组巢式引物进行扩增,腺病毒阳性样本6份,均为腺病毒3型;测序成功的5株核苷酸、蛋白质序列同源性均为100.0%。第3组巢式引物扩增、分析效果较好,阳性率为71.4%。结论成品试剂盒可进行不明原因发热疫情的病原筛查;多重巢式PCR扩增法可增加病毒检出率,对病毒分型和序列分析。Objective To enhance the laboratory capacity ability for rapid diagnosis of sudden fever epidemics;to improve and optimize pathogen detection methods.Methods A multiplex nested PCR amplification method was used to screen the pathogens of cases of a clustered fever.Meanwhile,the amplified products were sequenced and compared by BLAST.Serotyping was carried out,the amplification effect of different primers were compared.Results A total of 7 swab samples were collected,which were screened by multiple respiratory pathogen nucleic acid inspection kit for 14 kinds of pathogens,among which 4 samples were positive for adenovirus,the positive rate was 57.1%.3 sets of nested primers were used for amplification,6 samples were positive for adenovirus,all were adenovirus type 3.5 strains were sequenced successfully,their nucleotides and protein sequences were 100.0%homologous.Using the 3 rd set of primers delivered best amplification and analysis results,the positive rate was 71.4%.Conclusion The commercial diagnosis kit can be used to screen pathogen of fever epidemic cause by unknown origins.While multiple nested PCR amplification method can increase the detection rate,perform genotyping and sequence analysis of virus.

关 键 词：呼吸道感染 腺病毒 基因分型 多重巢式PCR 

分 类 号：R117[医药卫生—公共卫生与预防医学]