Virion RNA was transcribed into complementary DNA (cDNA) by reverse transcriptase, cDNA was amplified by polymerase chain reaction (PCR) and the products of PCR were purified. RNA sequence analysis was performed by the dideoxynucleotide chain termination method using synthetic oligodeoxynucleotide primers. Phylogenetic analysis of the sequencing data was performed with Megalign (version 1.03) and Editseq (version 3.69) software. Since 1995, there have been 3 different genetic lineages of influenza A (H1N1) virus HA1 gene co-circulating in men in Shenzhen city, China. The addition and deletion of one of the potential glycosylation sites at 54 and 155 positions of amino acid sequences on HA1 protein domain of H1N1 viruses isolated recently was found compared with those of A/Singapore/6/86 (H1N1) virus. Some differences in amino acid sequences on HA1 protein molecules occurred among H1N1 viruses tested and A/Singapore/6/86 (H1N1) virus. It is concluded that the occurrence of influenza A (H1N1) activity 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 the HA1 protein domains and due to the occurrence of influenza A (H1N1) virus with an “O” phase feature.
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