2007年至2008年昆明地区腹泻患儿轮状病毒非结构蛋白基因特征的分析

目的 探讨轮状病毒非结构蛋白(NSP4)基因的特征及变异情况.方法 在2007年和2008年昆明医学院第一附属医院儿科住院的96例秋冬季腹泻患儿中,随机选取轮状病毒阳性的12例(其中2007年6例,2008年6例)患儿粪便标本,采用RT-PCR扩增NSP4基因,并进行测序,测序结果 用DNAassist软件对核苷酸序列进行翻译,并用Clustal-mp软件与Genbank Datebase中的4株人RV(Wa、KUN、AU-1、Hochi)、3株动物RV(EW、OSU、SAll)和中国北京、上海、广州的流行株的NSP4序列进行分析比较.结果 (1)选取的2007年6份和2008年6份样本均可以扩增出NSP4的全长片段,测序表明:2007年6份样本中有5株属于Wa株,1株属于KUN株;2008年6份样本中也有5株属于Wa株,1株属于KUN株.10株Wa株与来自GenBank Database的4株人RV(wa、KUN、AU-1、Hochi)和3株动物RV(EW、OSU、SAll)同源性分别为96.3%、85.3%、84.7%、95.1%和57.7%、92%、82.8%,与中国北京、上海、广州的流行株同源性均在95%以上;2株KUN株与以上来自GenBank Database的标准株的同源性分别为85.3%、98.2%、82.8%、85.9%、60.1%、85.3%、94.3%,和其他地区流行株的同源性为85%左右; (2)以Wa株为参考,10株Wa株的NSP4基因变异发生在第34位氨基酸,第76位氨基酸,第141位氨基酸,第145位氨基酸,第161位氨基酸和第169位氨基酸;以KUN株为参考,2株KUN株的NSP4基因变异仅在第87位氨基酸和第140位氨基酸.结论 2007年和2008年昆明地区流行株NSP4基因为Wa组和KUN组,以Wa组为主.

Analysis on Genetic Characteristics of the Nonstructural Protein NSP4 of Human Rotavirus Strains in Infants with Diarrher in 2007 and 2008 in Kunming

Objective To study the genetic characteristics and genetic variation of the nonstructural protein NSP4 of Human rotavirus strains.Methods We selected twelve positive RV samples among 96 infants with diarrhea who were hospitalized in the Department of Peadiatrics,the First Affiliated Hospital of Kunming Medical University from October to December in 2007 and 2008.NSP4 genes was amplified by RT-PCR and then cDNAs were sequenced.The homology of the nucleotide sequences of the NSP4 genes were analyzed by DNA assist and Clustal-mp.Results(1)NSP4 gene was amplifie from 12(6 of 2007 and 6 of 2008)RV positive samples.Among the twelve Kunming strains,the nucleotide homologies of the ten Wa strains to the four HRV standard strains(Wa,KUN,AU-1 and Hochi)and three animal RV(EW,OSU and All)were 96.3%,85.3%,84.7%,95.1% and 57.7%,92%,82.8%.Compared with the epidemic strains in other different areas of China,the nucleotide homology of the NSP4 gene was over 95%.Compared with the standard strains from the above GenBank Database,the nucleotide homology of the two KUN strain were 85.3%,98.2%,82.8%,85.9% and 60.1%,85.3%,94.3%.And compared with the epidemic strains other different areas of China,the nucleotide homology of the NSP4 gene was about 85%.(2)6 unique conserved amino acids of 10 Wa strains differing from Wa at aa34,aa76,aa141,aa161 and aa169 were identified within NSP4;2 unique conserved amino acids of 2 KUN strains differing from KUN at aa87 and aa140 were identified within NSP4.Conclusions Among the NSP4 of the twelve Kunming strains,the genotypes are Wa strain and KUN strain.The most dominant genotype of NSP4 is Wa strain.