Ou Xin-Hua, Zhang Ru-Sheng, Song Ke-Yun, Chen Tian-Mu, Liu Ru-Chun, Sun Bian-Cheng, Chen Fa-Ming
Changsha Municipal Center for Disease Control and Prevention, Changsha 410001, China.
Bing Du Xue Bao. 2012 May;28(3):265-71.
In order to investigate the transmission risk of H5N1 avian influenza viruses (AIV) from sewage in Changsha poultry markets, the evolution relationship and molecular characteristics of non-structural (NS) genes of H5N1 AIV from sewage were analyzed. Nine H5N1 AIV environmental sewage specimens were collected from Changsha poultry markets. The NS genes were amplifyed by PCR and then sequenced with TA cloning. Amino acid(aa) sequence alignment and phylogenetic tree analysis were conducted by Lasergene and Mega5 software. Eight NS genes TA cloning were constructed successfully. Phylogenetic tree indicated that they were belonged to the allele A subgroup. Aa homology analysis showed 90.1% 92.5% identity in NS1 proteins and 91.0% - 92.6% identity in NS2 proteins compared with reference viruses of the allele A (A/chicken/ Hubei/ w h/ 1999). The homologies of the amino sequences of NS1 and NS2 in this study were 93.8%-100.0% and 98.4%-100.0%, respectively. The C terminal of all eight H5N1 NS1 proteins from sewage in poultry markets carried a ESEV of PL motif and the 92 amino acids were E, furthermore, the 80 to 84aa were missed which were the characteristics of highly pathogenic AIV. The NS genes of H5N1 AIV from sewage in poultry markets have molecular characteristics of highly pathogenic and have the potential risk of H5N1 virus spreading.
为调查长沙家禽市场污水中H5N1禽流感病毒（AIV）的传播风险，分析了污水中H5N1 AIV非结构（NS）基因的进化关系和分子特征。从长沙家禽市场采集了9份H5N1 AIV环境污水样本。通过PCR扩增NS基因，然后用TA克隆进行测序。利用Lasergene和Mega5软件进行氨基酸（aa）序列比对和系统发育树分析。成功构建了8个NS基因的TA克隆。系统发育树表明它们属于等位基因A亚组。与等位基因A（A/鸡/湖北/wh/1999）的参考病毒相比，氨基酸同源性分析显示NS1蛋白的同一性为90.1% - 92.5%，NS2蛋白的同一性为91.0% - 92.6%。本研究中NS1和NS2氨基酸序列的同源性分别为93.8% - 100.0%和98.4% - 100.0%。家禽市场污水中所有8个H5N1 NS1蛋白的C末端带有PL基序的ESEV，且第92位氨基酸为E，此外，缺失了第80至84位氨基酸，这是高致病性AIV的特征。家禽市场污水中H5N1 AIV的NS基因具有高致病性的分子特征，存在H5N1病毒传播的潜在风险。
