摘要
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．O％（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0300IN／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。
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 per- formed 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 posi- tive 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 patho- gen. Analysis among the 5 positve isolates based on the complete VP1 sequences showed 97.9% -99.5% ho- mology. Data from homologous comparisons indicated that these isolates had the highest nucleotide acid iden- tity 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 and2012, respectively. Phylogenetic tree in VP1 region showed that isolates of this outbreak belonged to geno- type D, the same clade with Changchun strain. Conclusion CVB5 was the major etiological agent correla- ted with this outbreak. The shift of predominant genotype might serve as one of the causes that associated with this outbreaks.
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2012年第7期610-613,共4页
Chinese Journal of Microbiology and Immunology
基金
河南省医学科技攻关项目（200702016）
河南省医学科技攻关计划省部共建项目（201201003）
