Huang Yong-Kun, Qi Qin, Hou Zong-Liu, Li Hai-Lin, Wen Ge-Sheng, Pang Wei, Zhou Li-Fang
Department of Pediatric, First Affiliated Hospital of Kunming Medical College, Kunming 650032, China.
Zhonghua Liu Xing Bing Xue Za Zhi. 2005 Dec;26(12):980-3.
To explore the molecular characteristics and molecular variation of human rotavirus (HRV) strains and to understand the relationship between clinical characteristics and epidemiology of different HRV-VP7 and NSP4.
Double-strand RNA of rotavirus extracted from stool samples was used as the template for reverse transcription of gene VP7, which was followed by nested PCR for VP7 typing. NSP4 genes from 22 epidemic strains of human rotavirus isolated in Kunming in 2002 and 2003 were amplified with RT-PCR. Then cDNAs were sequenced and compared with 4 human rotavirus NSP4 (Wa, KUN, AU-1, Hochi)) and 3 animal rotavirus NSP4 (EW, OSU, SA11) available in the GenBank while the epidemic strains of human rotavirus isolated in different areas of China were compared, using the Clustal-mp, DNAssist, MEGA2 software. The G serotype of VP7 was analysed by PCR.
Serotype G1 was prevalent in 2002 while serotype G3 was the prevalent in Kumming in 2003. The NSP4 genes from 22 epidemic strains of human rotavirus isolated in Kunming in 2002 and 2003 belonged to Wa with highly conservative amino acid. Samples isolated in the same years but not in the same area shared higher homology. Symptoms associated with heavy diarrhea did not seem to be associated with NSP4 molecular variation (P > 0.05).
Obvious variations of VP7 typing were seen in the same season, as well as in different areas and years. Due to the stable nature of NSP4, it seem to be a better candidate for vaccine production, than VP7.
探讨人轮状病毒(HRV)毒株的分子特征和分子变异情况,了解不同HRV-VP7和NSP4的临床特征与流行病学之间的关系。
以从粪便样本中提取的轮状病毒双链RNA为模板,反转录基因VP7,随后进行巢式PCR进行VP7分型。用RT-PCR扩增2002年和2003年在昆明分离的22株人轮状病毒流行株的NSP4基因。然后对cDNA进行测序,并与GenBank中可用的4种人轮状病毒NSP4(Wa、KUN、AU-1、Hochi)和3种动物轮状病毒NSP4(EW、OSU、SA11)进行比较,同时使用Clustal-mp、DNAssist、MEGA2软件比较在中国不同地区分离的人轮状病毒流行株。通过PCR分析VP7的G血清型。
2002年G1血清型流行,而2003年G3血清型在昆明流行。2002年和2003年在昆明分离的22株人轮状病毒流行株的NSP4基因属于Wa型,氨基酸高度保守。同年但不同地区分离的样本具有较高的同源性。与严重腹泻相关的症状似乎与NSP4分子变异无关(P>0.05)。
在同一季节、不同地区和年份均观察到VP7分型有明显变异。由于NSP4性质稳定,它似乎比VP7更适合作为疫苗生产的候选对象。