2010-2012年广州地区人冠状病毒分子进化分析

Molecular evolution of human coronavirus in Guangzhou from 2010 to 2012

目的 了解2010-2012年广州地区发热呼吸道感染患者的人冠状病毒各亚型分子进化情况.方法 采用RT-PCR法扩增HCoV-OC43、HCoV-229E和HCoV-NL63阳性标本的NP、RdRp和S基因部分片段,构建载体后进行测序,利用Bio-edit、Mega4.0、Clustal1.83等生物信息学软件对测序成功的基因序列进行比对和分析,建立HCoV-OC43、HCoV-229E和HCoV-NL63不同基因区域的分子进化树.结果 2010-2012年期间,HCoV-OC43、HCoV-229E和HCoV-NL63亚型在广州均未检测到重大变异株或重组株.HCoV-OC43与Belgium和中国香港来源流行株亲缘关系较近（GenBank登录号JN129834和AY903460）;HCoV-229E与Amsterdam来源的病毒株亲缘关系最近（GenBank登录号JX503060）;HCoV-NL63与中国北京和Amsterdam来源的流行株亲缘关系较近（GenBank登录号JX104161和DQ445911）.各亚型NP和RDRp为保守基因,S基因变异较大.结论 2010-2012年期间,广州至少存在3种HCoV-OC43、HCoV-229E和HCoV-NL63流行株,没有发现重大变异株或重组株.各亚型NP和RDRp均为保守基因,可用于病毒的检测,S基因变异较大,适用于对分子变异规律的研究.

Objective To investigate the molecular evolution characteristics of human coronavirus （HCoV） subtypes in patients with fever and respiratory tract infection in Guangzhou from 2010 to 2012.Methods Partial fragments of NP,RdRp and S genes of HCoV-OC43,HCoV-229E and HCoV-NL63 positive samples were amplified by RT-PCR and sequencing.Bioinformatics software,including Bio-edit,Mega4.0 and Clustal1.83 were used for comparison and analysis of NP,RDRp and S gene sequences.Molecular evolutionary tree of different gene regions of HCoV-OC43,HCoV-229E and HCoV-NL63 were built.Results No remarkable variation or recombinant strain of HCoV-OC43,HCoV-229E and HCoV-NL63 was found in Guangzhou during 2010-2012.The HCoV-OC43 substrains were genetically closest to the strains found in Belgium and Hong Kong （GenBank accession number JN129834 and AY903460）.HCoV-229E substrains were genetically closest to those found in Amsterdam （GenBank accession number JX503060） and HCoV-NL63 most genetically close to those in Amsterdam and Beijing （GenBank accession number JX104161 and DQ445911）.The NP and RDRp genes of all subtypes were highly conserved,while S gene was more variable.Conclusions There were at least 3 substrains of HCoV-OC43,HCoV-229E and HCoV-NL63 epidemic in Guangzhou during 2010-2012,and no remarkable variation or recombinant viral strain was found.The NP and RDRp genes of all subtypes were highly conserved and can be used in virus detection,while S gene was more variable and suitable for phylogenetic and variation study.