CAJ | 학술논문

目的鉴定2011年引起河南省漯河地区一起病毒性脑炎(VE)暴发的病原体,并对分离的柯萨奇病毒B5(CVB5)进行基因特征分析.方法采集漯河地区病毒性脑炎暴发期间29例患者的5份咽拭子、21份粪便和14份脑脊液,采用实时荧光RT-PCR方法分别检测总肠道病毒(PE)、EV71及CA16病毒核酸.所有标本均进行病毒分离培养,对其中5例患者的2份粪便和3份脑脊液的阳性分离产物进行VP1和5′-UTR区核苷酸序列测定及亲缘进化分析.结果实时荧光RT-PCR结果显示,所有临床标本EV71和CA16病毒核酸检测均为阴性,咽拭子、粪便和脑脊液的PE核酸检测阳性率分别为60.0％(3/5)、61.9％(13/21)和85.7％ (12/14).咽拭子、粪便和脑脊液病毒分离率分别为20.0％(1/5)、25.0％(5/21)和29.0％ (4/14).VP1和5′-UTR区核苷酸序列BLAST分析及分子分型结果显示为CVB5,对其中5株分离毒株进行VP1全长基因分析显示彼此之间核苷酸同源性为97.9％～ 99.5％.与其同源性最近的毒株是2010年引起长春手足口病暴发的CVB5分离株CC10/10/Changchun,核苷酸同源性为97.1％～98.1％.与2010及2012年河南省平顶山地区的脑炎分离株COXB5/Henan/2010和03001N/HN/CHN/2011/CB5的同源性分别为89.0％～89.6％和91.8％～92.5％.VP1区遗传进化树显示此次分离株为基因型D,且与长春株成一簇.结论导致此次病毒性脑炎暴发的病原体为肠道病毒CVB5,优势基因型的变迁可能与此次暴发相关.
목적 감정2011년인기하남성탑하지구일기병독성뇌염(VE)폭발적병원체,병대분리적가살기병독B5(CVB5)진행기인특정분석.방법 채집탑하지구병독성뇌염폭발기간29례환자적5빈인식자、21빈분편화14빈뇌척액,채용실시형광RT-PCR방법분별검측총장도병독(PE)、EV71급CA16병독핵산.소유표본균진행병독분리배양,대기중5례환자적2빈분편화3빈뇌척액적양성분리산물진행VP1화5′-UTR구핵감산서렬측정급친연진화분석.결과 실시형광RT-PCR결과현시,소유림상표본EV71화CA16병독핵산검측균위음성,인식자、분편화뇌척액적PE핵산검측양성솔분별위60.0％(3/5)、61.9％(13/21)화85.7％ (12/14).인식자、분편화뇌척액병독분리솔분별위20.0％(1/5)、25.0％(5/21)화29.0％ (4/14).VP1화5′-UTR구핵감산서렬BLAST분석급분자분형결과현시위CVB5,대기중5주분리독주진행VP1전장기인분석현시피차지간핵감산동원성위97.9％～ 99.5％.여기동원성최근적독주시2010년인기장춘수족구병폭발적CVB5분리주CC10/10/Changchun,핵감산동원성위97.1％～98.1％.여2010급2012년하남성평정산지구적뇌염분리주COXB5/Henan/2010화03001N/HN/CHN/2011/CB5적동원성분별위89.0％～89.6％화91.8％～92.5％.VP1구유전진화수현시차차분리주위기인형D,차여장춘주성일족.결론 도치차차병독성뇌염폭발적병원체위장도병독CVB5,우세기인형적변천가능여차차폭발상관.
Objective To identify the pathogen that caused an outbreak of viral encephalitis in Henan area in 2011.Phylogenic analysis was carried out on Coxsackie-virus B5 (CVB5) which was isolated during this outbreak.Methods Five throat swab,21 stool and 14 cerebrospinal fluid (CSF) specimens were collected from 29 inpatients during this outbreak.Viral isolation and real time RT-PCR were then performed for all specimens.Viral nucleic acid of enterovirus 71 (EV71),coxsackievirus A 16 (CA16) and pan-enterovirus (PE)were detected by real time RT-PCR.Phylogenetic tree based on entire VP1 sequences was constructed among CVB5 isolates from 2 stool and 3 CSF specimens of 5 inpatients and others published data retrieved from GenBank.Results The real time RT-PCR results showed that the PE nucleic acid positive rates of throat swab,stool and CSF specimens were 60.0％ (3/5),61.9％ (13/21) and 85.7％ (12/14) respectively.All of these specimens were negative for EV71 and CA16.The isolation rates of throat swab,stool and CSF specimens were 20.0％ (1/5),25.0％ (5/21) and 29.0％ (4/14),respectively.BLAST with both VP1 and 5′-UTR sequences and molecular typing indicated that CVB5 was the main pathogen.Analysis among the 5 positve isolates based on the complete VP1 sequences showed 97.9％-99.5％ homology.Data from homologous comparisons indicated that these isolates had the highest nucleotide acid identity with the Changchun CVB5 CC10/10/Changchun strain (97.1％-98.1％) which caused hand,foot,and mouth disease (HFMD) outbreak in Changchun in 2010,and lower identity (89.0％-89.6％ and 91.8％-92.5％) with the COXB5/Henan/2010 and 03001N strain isolated from Pingdingshan,Henan in 2010 and 2012,respectively.Phylogenetic tree in VP1 region showed that isolates of this outbreak belonged to genotype D,the same clade with Changchun strain.Conclusion CVB5 was the major etiological agent correlated with this outbreak.The shift of predominant genotype might serve as one of the causes that associated with this outbreaks.