Wu Shao-Hui, Shu Yue-Long, Zhao Zhuo, Yao Wen-Qing, Yu Wei, Zhang Mei-Mei, Cui Jian-Qiu, Liu Min, Fu Rong-Hua, Zhao Xiao-Guang
Liaoning Center for Disease Control & Prevention, Shenyang 110005, China.
Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi. 2009 Jun;23(3):174-6.
To understand the HA1 genetic variation characterization of influenza virus subtype H3N2 circulated from 2001 to 2006 in Liaoning local area.
Viral RNA was extracted and transcribed into cDNA by reverse transcriptase and amplified by PCR. The product of PCR was purified by QIAgen purification kits,and sequenced by ABI 3100avant. The sequence data were analyzed phylogenetically by Sequence software with epidemic records. Finally, the phylogenetic trees were drawn according to deduced amino acid sequences of influenza virus H3N2 from 2000 to 2006 in the NCBI database.
The seven HA1domain sequences of H3N2 influenza viruses circulated from 2001 to 2006 in Liaoning local area had been analyzed. Compared with WHO 2004-2006 H3N2 vaccine A/California/7/2004, 12 bases had changed, 4 positions had amino acid substitution in 62 * > E, 182 T > 1,224 S > A,225 C > Y. 224 and 225 are RBS (Receptor binding site). The homology is lower than 98%. Phylogenetic tree showed Liaoning H3N2 2006 strains and Zhejiang 2005 strains were similar to WHO Northern hemisphere winter 2006-2007 Vaccine A/Wisconsin/67/2005 (H3N2)-like virus and grouped together to form an independent cluster even though several bases were still different.
The HA1 domain of HA gene of influenza viruses (H3N2) isolated from 2001-2006 in Liaoning local area showed base mutation, amino acid sequence difference compared to A/California/7/2004 (2005-2006 vaccine), suggesting it might be the main cause leading to the spread of influenza. The sequence analysis showed Liaoning 2006 H3N2 strains were similar to those from Southern area which suggested that further surveullance should be conducted to monitor the virus mutation in circulation.
了解2001年至2006年在辽宁地区流行的H3N2亚型流感病毒的HA1基因变异特征。
提取病毒RNA,通过逆转录酶逆转录成cDNA,用PCR扩增。PCR产物用QIAgen纯化试剂盒纯化,并用ABI 3100avant测序。利用Sequence软件结合流行记录对序列数据进行系统发育分析。最后,根据NCBI数据库中2000年至2006年流感病毒H3N2的推导氨基酸序列绘制系统发育树。
分析了2001年至2006年在辽宁地区流行的H3N2流感病毒的7个HA1结构域序列。与WHO 2004 - 2006 H3N2疫苗株A/California/7/2004相比,有12个碱基发生了变化,62位* > E、182位T > I、224位S > A、225位C > Y这4个位置有氨基酸替换。224位和225位是受体结合位点。同源性低于98%。系统发育树显示,辽宁2006年的毒株和浙江2005年的毒株与WHO 2006 - 2007年北半球冬季疫苗株A/Wisconsin/67/2005 (H3N2) - like病毒相似,尽管仍有几个碱基不同,但它们聚集在一起形成一个独立的簇。
2001 - 2006年在辽宁地区分离的流感病毒(H3N2)的HA基因的HA1结构域与A/California/7/2004(2005 - 2006年疫苗株)相比,显示出碱基突变和氨基酸序列差异,提示这可能是导致流感传播的主要原因。序列分析表明,辽宁2006年的H3N2毒株与南方地区的毒株相似,提示应进一步开展监测以监测流行中的病毒变异。