Zhang Ye, Wen Le-ying, Li Zi, Guo Jun-feng, Wang Min, Guo Yuan-ji
National Institute of Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100052, China.
Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi. 2004 Mar;18(1):16-9.
To understand the antigenic and genetic characteristics of influenza A H3N2 viruses circulated in man in China from 2000 to 2002.
Embryonated chicken eggs inoculated with virus for amplification of viral yield. The harvesting egg allantoic fluids with influenza viruses were provided for testing antigen and RNA extraction. Virion RNA was transcribed into cDNA by reverse transcriptase, cDNA amplified by PCR, and the product of PCR was purified. Afterward RNA sequence analysis was performed by the dideoxynucleotide chain termination method using synthetic oligodeoxynucleotide primers. Finally the phylogenetic tree was analyzed with MegAlign software.
The H3N2 viruses isolated during 2000-2002 were different in amino acid sequences on HA1 domain protein molecule from those of A/Wuhan359/1995 H3N2 as well as those of A/Sydney/7/1997 H3N2 strains. There were four different positions of amino acid sequence on HA1 domain protein molecule among the H3N2 viruses isolated in 2000 and during 2001-2002. They located at 83, 186, 202, 222 and 225 position, respectively. Of them 83 and 186 were in antigenic determinant E and B, respectively. The others located at left wall of the receptor binding site (RBS).
From the end of 2001 to the beginning of 2002, the influenza epidemic in Northern China caused by H3N2 virus was due to occurrence of antigenic and genetic changes of influenza A(H3N2) virus.
了解2000年至2002年在中国人群中流行的甲型H3N2流感病毒的抗原性和基因特征。
接种病毒于鸡胚中以扩增病毒产量。收集含流感病毒的鸡胚尿囊液用于抗原检测和RNA提取。病毒粒子RNA通过逆转录酶转录为cDNA,cDNA经PCR扩增,PCR产物纯化。随后使用合成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软件分析系统发育树。
2000 - 2002年分离的H3N2病毒在HA1结构域蛋白分子上的氨基酸序列与A/武汉359/1995 H3N2以及A/悉尼/7/1997 H3N2毒株不同。2000年分离的H3N2病毒与2001 - 2002年分离的H3N2病毒在HA1结构域蛋白分子上有四个不同的氨基酸序列位置。它们分别位于第83、186、202、222和225位。其中83和186分别位于抗原决定簇E和B中。其他位于受体结合位点(RBS)的左壁。
2001年底至2002年初,中国北方由H3N2病毒引起的流感流行是由于甲型(H3N2)流感病毒发生了抗原性和基因变化。