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目的 了解一株再次从流感患儿中分离出禽H9N2流感毒株的基因组特性 ,并弄清它的来源。方法 病毒在鸡胚中传代 ,从收获的尿囊液中提取RNA ,通过逆转录合成cDNA ,cDNA用PCR扩增。PCR产物用纯化试剂盒纯化 ,接着做核苷酸序列测定 ,然后用MegAlign(Version 1 0 3)和Editseg(Version 3 69)软件进行基因进化树分析。结果 A 广州 333 99(H9N2 )毒株的基因组属于禽流感病毒 ,但它明显不同于A Duck HongKong Y439 97毒株。同时不含有任何人流感病毒基因节段 ,其基因组中有 4个基因节段 (分别编码HA、NA、NP和NS蛋白 )来自G9毒株基因系 ,而其余 4个基因节段 (分别编码PB2、PB1、PA和M蛋白 )来自G1毒株基因系。结论 A 广州 333 99(H9N2 )病毒是G9和G1毒株通过基因重配而来的重配株 ,它最大可能性直接来自禽。进一步证实了禽H9N2毒株能感染人 ,同时首次证实了H9N2不同基因系毒株间 ,在自然界中也能发生基因重配
OBJECTIVE: To understand the genomic characteristics of a strain of H9N2 strain isolated from infants once again and to clarify its origin. Methods The virus was passaged in chicken embryos, RNA was extracted from the harvested allantoic fluid, cDNA was synthesized by reverse transcription, and cDNA was amplified by PCR. The PCR product was purified using a purification kit followed by nucleotide sequencing followed by phylogenetic tree analysis using MegAlign (Version 1 0 3) and Editseg (Version 3 69) software. Results A The genome of the Guangzhou 333 99 (H9N2) strain belongs to the avian influenza virus, but it is significantly different from the strain A Duck HongKong Y43997. And does not contain any human influenza virus gene segments. Four gene segments (HA, NA, NP and NS proteins, respectively) are contained in the genome from the G9 strain gene line while the remaining four gene segments (encoding PB2 , PB1, PA and M proteins) are from the G1 strain gene line. Conclusion A Guangzhou 333 99 (H9N2) virus is a reassortant of G9 and G1 strains obtained by genetic recombination. Its greatest possibility is directly from the poultry. Further confirmed that the bird H9N2 strain can infect humans, at the same time for the first time confirmed that H9N2 different strains of genes between strains in the nature of genetic reassignment