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对2005年11月8日安徽省铜陵市人民医院报告的一例孕妇不明原因肺炎病例的死亡病因进行研究。采集病人的气管吸出物及血液标本,用RT-PCR和Real-ti me PCR方法检测流感病毒A/M、A/H5N1、A/H7N7、A/H9N1亚型特异性核苷酸片段;气管吸出物接种SPF鸡胚进行病毒分离,并对分离物进行鉴定和序列测定及分析;利用血凝抑制试验检测血清标本抗体。结果表明病人气管吸出物可以检测到甲型流感病毒M片段及H5亚型的特异性HA基因。2005年11月9日采集的血清标本用Real-ti me PCR检测到甲型流感病毒M基因。从病人的气管吸出物中分离到H5N1病毒(A/Anhui/1/2005),对病毒的HA基因序列结果进行分析表明病毒是禽源的,其主要依据是受体结合位点第226~228位氨基酸位点(QSG)为禽流感病毒所特异,HA受体连接肽仍为9个碱性氨基酸(LRERRRKRP);基因进化树分析显示,HA基因与禽源病毒进化距离接近。发病后7、8、9d的血清H5N1禽流感病毒HI抗体小于20。对该病例的病原学研究证明,该病例为H5N1禽流感感染病例。
On November 8, 2005, Tongling City People’s Hospital reported that a case of unexplained pneumonia in pregnant women, the cause of death were studied. The tracheal aspirates and blood samples of patients were collected. The specific nucleotide fragments of A / M, A / H5N1, A / H7N7 and A / H9N1 subtypes were detected by RT-PCR and Real- SPF chicken embryos were inoculated for virus isolation, and the isolates were identified and sequenced and analyzed. The serum samples were detected by hemagglutination inhibition test. The results show that patients with tracheal aspirates can detect influenza A virus M fragment and H5 subtype specific HA gene. Serum samples collected on November 9, 2005 were tested for Influenza A virus M gene by Real-ti me PCR. The H5N1 virus (A / Anhui / 1/2005) was isolated from the tracheal aspirate of the patient and the results of the HA gene sequence analysis of the virus indicated that the virus was of poultry origin and was mainly based on receptor binding sites 226-228 The amino acid site (QSG) was specific to the avian influenza virus, and the HA receptor linker was still 9 basic amino acids (LRERRRKRP). The phylogenetic tree analysis showed that the HA gene was evolved closely to the avian virus. Serum H5N1 avian influenza virus HI antibody was less than 20 at 7, 8 and 9 days after onset. Etiological studies of the case proved that the case of H5N1 bird flu infection.