2016年福建省输入性黄热病的分子诊断

Molecular diagnosis of yellow fever imported into Fujian province, 2016

目的 应用分子检测方法,对输入性黄热病病例进行实验诊断.方法 采集病例血清和尿液标本,应用实时荧光定量逆转录PCR（Real-time RT-PCR）法检测黄热病毒特异性核酸;套式RT-PCR法扩增病毒基因片段,序列测定或片段长度多态性法鉴别野毒株或疫苗株.结果 Real-time RT-PCR法检测5例黄热病毒核酸阳性病例,病毒在尿液标本中存在时间较血清中存在时间更长,序列测定及限制性片段长度多态性法证实Angola71型病毒感染.结论 同时采用血液和尿液检测可以提高黄热病检测敏感性,RFLP分析方法可快速鉴别野病毒感染,排除因疫苗接种后可能出现的阳性结果.

Objective To make laboratorial diagnosis of imported yellow fever （YF） cases in Fujian province with molecular method.Methods Serum and urine samples were collected from suspected cases at various time-points post illness onset.Real-time RT-PCR and nested RT-PCR were performed respectively for viral specific nucleotide detection and fragment amplification.Sequencing and restrictive fragment length polymorphism （RFLP） method were used to identify the wild virus infection.Results A total of five cases with wild yellow fever virus （YFV） infection were confirmed in this study.It revealed that the viral agent belonged to Angola-71 like YFV,and the duration of viral agent in urine was longer than that in serum.Conclusions Simultaneous detection of serum and urine samples would increase detection sensitivity,and further RFLP method contributed to rapid identification of wild YFV infection and exclusion of positive result due to recent vaccination.