CAJ | 학술논문

目的了解2011年上海地区手足口病重症和轻症患儿中肠道病毒71型(EV71)分离株VP1、VP4区的基因特征.方法对来自2011年重症与轻症手足口病患儿的各5株EV71分离株进行VP1、VP4全序列的RT-PCR扩增测序,并与美国国立生物技术信息中心公布的EV71 A、B、C基因型代表株进行核苷酸、氨基酸比对分析和系统进化分析.结果轻、重症患儿的EV71分离株之间VP1基因的核苷酸同源性为96.0％～98.1％;VP4基因的核苷酸同源性为93.7％～99.5％.轻、重症患儿的EV71分离株与C基因型代表株比较接近,VP1区核苷酸同源性分别为86.9％～98.2％、87.4％o～98.5％,VP4区核苷酸同源性分别为85.5％～100.0％、84.5％～99.5％,其中与2008年安徽省阜阳市的EV71流行株(C4亚型)VP1区核苷酸同源性分别可达97.0％～98.2％、97.9％～98.5％,VP4区核苷酸同源性分别可达96.1％～100.0％、97.1％～99.5％.3例重症患儿分离株在VP1和VP4的天冬酰胺(N)282丝氨酸(S)、苏氨酸(T)7丙氨酸(A)同时发生变异.结论 2011年上海地区10例轻、重症手足口病患儿中分离的EV71流行株均属C基因型的C4亚型；3例重症患儿分离株在VP1和VP4的N282S、T7A同时发生变异.
목적 료해2011년상해지구수족구병중증화경증환인중장도병독71형(EV71)분리주VP1、VP4구적기인특정.방법 대래자2011년중증여경증수족구병환인적각5주EV71분리주진행VP1、VP4전서렬적RT-PCR확증측서,병여미국국립생물기술신식중심공포적EV71 A、B、C기인형대표주진행핵감산、안기산비대분석화계통진화분석.결과 경、중증환인적EV71분리주지간VP1기인적핵감산동원성위96.0％～98.1％;VP4기인적핵감산동원성위93.7％～99.5％.경、중증환인적EV71분리주여C기인형대표주비교접근,VP1구핵감산동원성분별위86.9％～98.2％、87.4％o～98.5％,VP4구핵감산동원성분별위85.5％～100.0％、84.5％～99.5％,기중여2008년안휘성부양시적EV71류행주(C4아형)VP1구핵감산동원성분별가체97.0％～98.2％、97.9％～98.5％,VP4구핵감산동원성분별가체96.1％～100.0％、97.1％～99.5％.3례중증환인분리주재VP1화VP4적천동선알(N)282사안산(S)、소안산(T)7병안산(A)동시발생변이.결론 2011년상해지구10례경、중증수족구병환인중분리적EV71류행주균속C기인형적C4아형；3례중증환인분리주재VP1화VP4적N282S、T7A동시발생변이.
Objective To analyze the VP1 and VP4 genetic region of enterovirus 71 (EV71)isolated from severe cases and mild cases with hanD-foot-mouth disease (HFMD) in Shanghai in 2011.Methods Five EV71 strains isolated from severe cases and five EV71 strains from mild cases in 2011 were included.Reverse transcription-polymerase chain reaction (RT-PCR) method was used to amplify and sequence the VP1 and VP4 genes of EV71,and then the sequencing results were compared with those of A,B,C genotype reference EV71 strains from GenBank by nucleotide alignment,amino acid alignment and phylogenetic tree analyses.Results The homogeneity between EV71 strains from severe cases and mild cases was 96.0％-98.1％ and 93.7％-99.5％ for VP1 and VP4 nucleotide sequences,respectively.The VP1 nucleotide sequences of 5 strains isolated from severe cases and 5 strains from mild cases in Shanghai shared 86.9％-98.2％ and 87.4％ 98.5％ identity with genotype C,respectively,while the homogeneity of VP4 nucleotide sequence was 85.5％-100.0％and 84.5％-99.5％,respectively.In addition,compared with the Fuyang EV71 strains (representative of C4 subtype),the strains from mild and severe cases shared homogeneity of 97.0％-98.2％ and 97.9％-98.5％ for VP1 gene,respectively,96.1％-100.0％ and 97.1％-99.5％ for VP4 gene,respectively.Among 3 strains isolated from severe cases,mutations at the residue 282 in the VP1 protein (N→S) and residue 7 in the VP4 protein (T→A) were discovered simultaneously.Conclusions The 10 EV71 strains isolated from severe and mild cases in Shanghai belong to subgenogroup C4.Among 3 strains isolated from severe cases,mutations at the residue 282 in the VP1 protein (N→S) and residue 7 in the VP4 protein(T→A) are discovered simultaneously.