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目的研究腹泻患儿的轮状病毒(RV)流行株非结构蛋白4(NSP4)的基因变异与临床的关系。方法对昆明地区2002年和2003年流行期不同程度腹泻患儿中分离出的22株轮状病毒流行株的NSP4基因进行RT-PCR扩增,并进行cDNA序列分析,与来自GenBank Database的4株人RV(Wa、KUN、AU-1、Hochi)和3株动物RV(EW、OSU、SAll)以及我国不同地区RV流行株NSP4的基因变异情况进行比较。结果氨基酸同源性表明昆明地区轮状病毒流行株间的同源性高达98.9%~99.4%,22株流行株全都属于Wa组。氨基酸进化树提示在Wa组内又包括3个亚组。NSP4基因变异与轮状病毒腹泻临床症状严重程度不相关(P>0.05)。结论2002年和2003年昆明地区轮状病毒流行株皆属Wa组,且其氨基酸序列极为保守;NSP4基因变异与轮状病毒腹泻临床症状严重程度不相关,但与流行发病的关系需继续研究。
Objective To study the relationship between the gene mutation of non-structural protein 4 (NSP4) of rotavirus (RV) epidemic children and clinical manifestations in children with diarrhea. Methods The NSP4 gene of 22 strains of rotavirus isolated from children with diarrhea in different stages of epidemic period in Kunming in 2002 and 2003 were amplified by RT-PCR and analyzed by cDNA sequence. The gene variants of human RV (Wa, KUN, AU-1, Hochi) and three animals RV (EW, OSU, SAll) and RV epidemic strain NSP4 in different regions of China were compared. Results The amino acid homology indicated that the homology between rotavirus strains in Kunming was as high as 98.9% -99.4%. All the 22 strains belong to Wa group. The amino acid phylogenetic tree suggests that there are 3 subgroups in Wa group. The variation of NSP4 gene was not related to the severity of clinical symptoms of rotavirus diarrhea (P> 0.05). CONCLUSION: The rotavirus strains in Kunming in 2002 and 2003 belong to group Wa, and their amino acid sequences are very conservative. The variation of NSP4 gene is not related to the severity of clinical symptoms of rotavirus diarrhea, but the relationship with epidemic should be further studied.