山东省部分地区脑炎与非脑炎手足口病肠道病毒71型VP1区基因特征  

Analysis of the genetic characteristics of Enterovirus 71 in infants with hand,foot and mouth disease who also had or who did not have encephalitis in Shandong Province in 2008

作　　者：张乐海[1] 马丽霞[1] 王世富[1] 刘霞[1] 姜忠强[1] 于华凤[1] 

机构地区：[1]山东大学齐鲁儿童医院,山东济南250022

出　　处：《中国病原生物学杂志》2010年第8期567-571,共5页Journal of Pathogen Biology

摘　　要：目的采用逆转录-聚合酶链反应-序列分析方法,对脑炎与非脑炎手足口病患儿粪便和脑脊液样本中肠道病毒71型(EV71)部分VP1区进行核苷酸和氨基酸序列比对,探讨核苷酸或氨基酸变异与不同临床类型的关系。方法以临床诊断和基因诊断方法确定手足口病和EV71感染,用自行设计的2对引物,在EV71 VP1区扩增核苷酸,连接T载体后进行序列分析。所得序列连接后与EV71各亚型和安徽阜阳流行株核苷酸序列,通过DNAMAN和BI-OEDIT软件比对,分析其基因特征,并构建系统发生树。结果 6个不同临床类型手足口病病人核苷酸序列与A型、B型、C型的同源性分别为82.2%～82.6%、82.9%～84.1%和89.1%～95.1%,其中与C4亚型同源性最高,为94.8%～95.1%,与安徽阜阳地区流行株的同源性在98.0%～98.4%之间;与安徽阜阳株氨基酸序列比对,同源性在99.3%～99.6%之间,且均在563位由I(异亮氨酸)变为T(苏氨酸),仅有1例脑炎病人zxuan_VP1_2438-3260(脑脊液)在584位A(丙氨酸)变为G(甘氨酸),1例HFMD病人A06_B816064.VP1_2438-3227(粪便)在736位由W(色氨酸)变为C(半胱氨酸),余未出现特征性改变。结论神经源性肺水肿、脑炎与非脑炎手足口病患儿分离株EV71 VP1区核苷酸序列和推导的氨基酸序列无特征性差异。Objective Reverse transcription polymerase chain reaction（RT-PCR） was used to detect hand,foot and mouth disease（HFMD） in the feces and cerebrospinal fluid samples of infants who also had or who lacked encephalitis.Enterovirus 71（EV71） VP1 region nucleotide and amino acid sequences were compared to explore the relationship between nucleotide or amino acid variations and different clinical types.Methods Clinical diagnosis and genetic diagnosis were used to determine HFMD caused by EV71 infection.Two pairs of self-designed primers were then used to amplify the EV71 VP1 nucleotides and ligate them to a T vector for sequencing.These nucleotide sequences were compared to EV71 subtype nucleotide sequences,such as the Fuyang,Anhui strains,using BIOEDIT and DNAMAN software in order to analyze their genetic characteristics and construct a phylogenetic tree.Results Six different clinical types of nucleotide sequences from patients were compared to A,B,and C-type standard strains,revealing a homology of 82.2%-82.6%,82.9%-84.1%,and 89.1%-95.1%,respectively.Sequences from patients had the highest homology with the C4 subtype,94.8%-95.1%,and a homology of 99.3%-99.6% compared to the Fuyang,Anhui strain.In addition,amino acid sequences had a homology of 99.3%-99.6% with the Fuyang strain.The amino acids had changed from I（isoleucine） to T（threonine） at site 563.Amino acids had changed from A（Ala） to G（glycine） at site 584 in the sample of zxuan_VP1_2438-3260（with encephalitis） and from W（tryptophan） to C（cysteine） at site 736 in the sample of A06_B816064.VP1_2438-3227（without encephalitis）.Conclusion EV71 VP1 nucleotide sequence analysis revealed that there were no characteristic differences in nucleotides and amino acids in infants with HFMD who also had or who did not have encephalitis.

关 键 词：肠道病毒71型 手足口病 脑炎 核苷酸序列 

分 类 号：R373.2[医药卫生—病原生物学]