Cheng X, Li L, He J
Municipal Hygiene and Anti-epidemic Station of Shenzhen, Shenzhen 580020.
Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi. 1999 Dec;13(4):340-4.
To understand the molecular bases of intention of influenza virus activity and emergence of "O" phase of influenza A(H1N1) strains in human population in Shenzhen in recently years, and also the evolutionary characterization of influenza A(H1N1) HA1 gene.
Virion RNA was transcribed into cDNA by reverse transcriptase, cDNA was amplified by PCR, the products of PCR were purified. Afterward, RNA sequence analysis was performed by the dideoxynucleotide chain termination method using synthetic oligodeoxynucleotide primers. Finally, phylogenetic analysis of the sequencing data was performed with MegAlign (version 1.03) and Editseq (version 3.69) softwares.
Since 1995, there were three different genetic lineages of influenza A(H1N1) virus HA1 gene cocirculating in men in Shenzhen city. Adding one and deleting one of potential glycosylation sites at 54 and 155 positions of amino acid sequences on HA1 protein domain of H1N1 viruses isolated recently was found as compared with those of A/Singapore/6/86(H1N1) virus. Meanwhile, there were some differences of amino acid sequences on HA1 protein molecules among H1N1 viruses tested and A/Singapore/6/86 (H1N1) virus.
The intention of influenza A(H1N1) virus activity occurred since 1995 was due to emergence of substitution in amino acid sequences, especially the appearance of one addition and one deletion of potential glycosylation sites on their HA1 protein domains, and also was due to occurrence of influenza A(H1N1) virus with feature of "O" phase.
了解近年来深圳市人群中甲型H1N1流感病毒活性趋势及“O”相毒株出现的分子基础，以及甲型H1N1流感病毒HA1基因的进化特征。
通过逆转录酶将病毒粒子RNA转录为cDNA，用PCR扩增cDNA，纯化PCR产物。随后，使用合成的寡脱氧核苷酸引物，通过双脱氧核苷酸链终止法进行RNA序列分析。最后，用MegAlign（版本1.03）和Editseq（版本3.69）软件对测序数据进行系统发育分析。
自1995年以来，深圳市人群中甲型H1N1流感病毒HA1基因有三种不同的遗传谱系共同流行。与A/新加坡/6/86（H1N1）病毒相比，发现最近分离的H1N1病毒HA1蛋白结构域氨基酸序列的54和155位存在潜在糖基化位点的增加和减少。同时，所检测的H1N1病毒与A/新加坡/6/86（H1N1）病毒在HA1蛋白分子的氨基酸序列上存在一些差异。
1995年以来甲型H1N1流感病毒活性趋势的出现是由于氨基酸序列的替代，特别是其HA1蛋白结构域上潜在糖基化位点的增加和减少，也是由于具有“O”相特征的甲型H1N1流感病毒的出现。
