CAJ | 학술논문

目的快速鉴定2014年济南市一起聚集性病毒性脑炎的病原体,分析其流行病学及病原学特征.方法采用统一的个案调查表对住院患儿逐一进行个案调查.4例患儿均采集粪便和脑脊液标本.采用实时荧光定量(RT) PCR方法检测总肠道病毒(PE)、肠道病毒71型(EV71)、柯萨奇病毒A组(CVA) 16、10和6型病毒核酸;PE阳性标本采用巢式反转录PCR方法扩增5'-UTR区基因,测定核苷酸序列并进行同源性分析.结果本起聚集性病毒性脑炎疫情首发病例出现于2014年4月8日,呈人传人的传播模式.RT-PCR结果显示,所有标本PE核酸检测为阳性,而EV71、CVA16、CVA10和CVA6核酸检测均为阴性.5'-UTR区核苷酸序列在美国生物技术信息中心网站经碱基局部对准检索工具分析(http://blast.ncbi.nlm.nih.gov/Blast.cgi)显示与肠道病毒柯萨奇病毒B5(CVB5)同源性最高,彼此之间核苷酸同源性为99.2％～ 100.0％,与2010年河南株CVB5(HQ998851)的核苷酸同源性为96.7％～97.4％.4例患儿经积极的抗病毒、抗感染及相关对症支持治疗,均痊愈出院,无后遗症发生.结论本起聚集性病毒性脑炎病原体为CVB5,与2010年河南株CVB5核苷酸同源性最高.RT-PCR和巢式反转录PCR方法可快速、准确地鉴定肠道病毒引起的病毒性脑炎.
목적 쾌속감정2014년제남시일기취집성병독성뇌염적병원체,분석기류행병학급병원학특정.방법 채용통일적개안조사표대주원환인축일진행개안조사.4례환인균채집분편화뇌척액표본.채용실시형광정량(RT) PCR방법검측총장도병독(PE)、장도병독71형(EV71)、가살기병독A조(CVA) 16、10화6형병독핵산;PE양성표본채용소식반전록PCR방법확증5'-UTR구기인,측정핵감산서렬병진행동원성분석.결과 본기취집성병독성뇌염역정수발병례출현우2014년4월8일,정인전인적전파모식.RT-PCR결과현시,소유표본PE핵산검측위양성,이EV71、CVA16、CVA10화CVA6핵산검측균위음성.5'-UTR구핵감산서렬재미국생물기술신식중심망참경감기국부대준검색공구분석(http://blast.ncbi.nlm.nih.gov/Blast.cgi)현시여장도병독가살기병독B5(CVB5)동원성최고,피차지간핵감산동원성위99.2％～ 100.0％,여2010년하남주CVB5(HQ998851)적핵감산동원성위96.7％～97.4％.4례환인경적겁적항병독、항감염급상관대증지지치료,균전유출원,무후유증발생.결론 본기취집성병독성뇌염병원체위CVB5,여2010년하남주CVB5핵감산동원성최고.RT-PCR화소식반전록PCR방법가쾌속、준학지감정장도병독인기적병독성뇌염.
Obgective To rapidly identify the pathogen of a clustered viral encephalitis in Jinan,2014,and to analyze the epidemiological and etiological characteristics.Methods Uniform questionnaire was used to investigate each case.Both stool and cerebrospinal fluid specimens of 4 children were collected.At first,real-time fluorescence quantitative(RT) PCR was performed to detect the viral nucleic acid of pan-enterovirus (PE),enterovirus 71 (EV71),coxsackievirus A16 (CVA1 6),coxsackievirus A10 (CVA10) and coxsackievirus A6 (CVA6).Then nested reverse transcription-PCR based on 5'-UTR gene of enterovirus was conducted for the PE positive specimens.The PCR products were sequenced and homologous analysis were performed.Results The initial case appeared on the April 8,2014 in the from of clustered viral encephalitis and the transmission mode was person-to-person.The RT-PCR resuits showed that PE was positive for all the specimens,while others including EV71,CVA16,CVA10 and CVA6 were negative.Analysis based on 5'-UTR through Basic Local Alignment Search Tool (http://blast.ncbi.nim.nih.gov/ Blast.cgi) sequences indicated that coxsackievirus B5 (CVB5) was the main pathogen and 99.2％-130.0％ homology from each other.Data from homologous comparisons showed that these sequences had homology of 96.7％-97.4％with Henan CVB5 strain(HQ998851).These 4 children all recovered after the treatment with antiviral medications,anti-infective agents,and supportive therapy.Conclusions CVB5 was responsible for the clustered viral encephalitis in Jinan,2014 and it had high homology with Henan CVB5 strain.RT-PCR and nested reverse transcription-PCR are rapid and effective methods to identify viral encephalitis caused by enterovirus.