人巨细胞病毒地方分离株的鉴定及其UL83序列分析  

Identification and UL83 sequence analysis of human cytomegalovirus area isolates

作　　者：史永林[1] 陈敬贤[1] 王明丽[1] 

机构地区：[1]安徽医科大学微生物学教研室,合肥230032

出　　处：《安徽医科大学学报》2006年第6期612-616,共5页Acta Universitatis Medicinalis Anhui

基　　金：教育部科学技术研究重点项目(编号:01052);安徽省"十五"生物医药重大科技专项(编号:01303003);安徽省教育厅自然科学基金资助项目(2006KJ320B)

摘　　要：目的鉴定从合肥市某医院先天性脑瘫患儿尿标本中分离的4株人巨细胞病毒（HCMV）毒株并对其UL83基因序列进行分析，为HCMV感染的预防及pp65蛋白疫苗的研制提供参考依据。方法应用细胞培养法复苏病毒，间接免疫荧光法检测pp65蛋白表达，并用特异性引物对UL73基因进行PCR，扩增的产物经凝胶纯化后测序；序列结果运用DNAMAN软件和GenBank登录的3株代表性HCMV毒株进行核苷酸和氨基酸序列比对。结果HCMV AD169株与HCMV-1分离株的UL83基因核苷酸及编码氨基酸序列的同源性均为100％，与其余3株的同源性也达99．26％～99．69％；4株HCMV分离株与Merlin株、Towne株在相应基因片段核苷酸及氨基酸的同源性最低的分别为98．95％，99．06％。结论UL83序列分析结果表明，HCMV UL83编码pp65蛋白的优势抗原决定簇氨基酸序列无变化。Objective To identify 4 human cytomegalovirus（HCMV） strains isolated from the urine of congenital cerebral palsy children of a certain hospital of Hefei and analyze their UL83 nucleotide sequence, to provide theories reference for prevention of HCMV infection and development of CMV-pp65 vaccine. Methods Cell culture, indirect immunofluorescence assay and PCR were applied in the study. The UL83 DNA of the 4 HCMV isolates were amplified using specific primers. PCR products were purified and sequenced. DNAMAN software was used to do nucleotide and amino acid pairwise-alignment with 3 representative HCMV strains in NCBI database. Results The nucleotide sequence and amino acid homologies of the UL83 region both were 100% between HCMV AD169 strain and HCMV-1 isolate, 99.26% - 99.69% between the other 3 isolates and HCMV AD169 strain. The lowest homologies of the nucleotide sequence and amino acid were 98.95% , 99. 06% respectively between 4 isolates and HC- MV Merlin ,Towne strain. Conclusion Analysis of homology of nucleotide and amino acid identically suggests that the amino acid sequence of predominance epitope of HCMV UL83 encoded pp65 antigen is unchange.

关 键 词：巨细胞病毒/分离和提纯 生物学鉴定法/方法 序列分析 DNA 

分 类 号：R373.9[医药卫生—病原生物学] R394-3[医药卫生—基础医学]