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目的测定2003年苏北地区无菌性脑膜炎爆发的病毒分离株部分基因序列,了解该流行株的分子生物学特点及遗传变异规律。方法随机选取3株分离的病毒株,用能特异性扩增肠道病毒VP1区3’段序列的两对引物012/011、040/011进行逆转录聚合酶链反应,扩增产物经凝胶纯化后测序。将序列输入GenBank用BLAST程序进行核苷酸和氨基酸序列比对;选取部分肠道病毒序列,经CLUSTALX1.83对齐后,在PHYLIP3.5c中构建进化树,了解分离毒株Echo30的进化关系。结果3株病毒测序结果均为Echo30型肠道病毒。进化树分析显示分离的Echo30毒株与国外1999年和2000年分离株的亲缘关系最近;但3株分离株自成一簇,与国外毒株存在地区差别。结论利用VP1区3’段序列可以快速正确区分肠道病毒分离株的型别,分离株与国外同型流行株在比较区段的遗传变异规律类似。
Objective To determine the partial gene sequence of the virus isolated from aseptic meningitis in northern Jiangsu in 2003 and to understand the molecular biological characteristics and genetic variation of the epidemic strain. Methods Three isolates were randomly selected and used for reverse transcriptase polymerase chain reaction with two pairs of primers that can specifically amplify the 3 ’sequence of VP1 region of enterovirus. The amplified products were separated by gel After purification, sequencing. The sequence was input into GenBank. The BLAST program was used to compare the nucleotide and amino acid sequences. A partial sequence of enterovirus was selected and aligned with CLUSTALX1.83. The phylogenetic tree was constructed in PHYLIP3.5c to understand the evolutionary relationship of the isolated strain Echo30. Results The sequencing results of the three viruses were Echo30 type enterovirus. Phylogenetic tree analysis showed that the isolated Echo30 strain had the closest relationship with foreign isolates from 1999 and 2000. However, the three isolates were self-clustered and differed from other strains abroad. Conclusion The sequence of VP1 3 ’segment can be used to distinguish the type of enterovirus isolates quickly and correctly. The genetic variation of the isolates in the comparison zone with that of foreign isolates is similar.