Isolation and characterization of H3N2 influenza A virus from turkeys
Isolation and characterization of H3N2 influenza A virus from turkeys
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DOI：
10.1637/7288-101304r
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发表时间：
2005-06-01
期刊：
影响因子：
1.4
通讯作者：
Saif, YM
中科院分区：
文献类型：
--
作者：
Tang, Y;Lee, CW;Saif, YM
在俄亥俄州的一个农场中，5个34周龄的火鸡繁殖蛋鸡群在2550只鸡的单独的房子里，从2004年1月底到2月初，鸡蛋产量下降。将鸡群的气管拭子（n = 60）、泄殖腔拭子（n = 50）和恢复期血清（n = 110）提交实验室进行诊断。尝试在无特定病原体的鸡胚和Vero和MDCK细胞中分离病毒。使用琼脂凝胶免疫扩散、血凝试验、血凝抑制试验、病毒中和试验、逆转录-聚合酶链反应、测序和系统发育分析进行病毒表征。一种假定的流感病毒在MDCK细胞中第一次传代时成功繁殖和分离，但最初不在Vero细胞或无特定病原体的鸡胚中。在MDCK细胞中传代两代后，该分离物可以在无特异性病原体的鸡胚中繁殖。分离病毒的初步序列分析证实，它是甲型流感病毒，与猪甲型流感病毒A/Swine/Illinois/100084/01（H1 N2）的基质基因几乎100%（235/236）相同。然而，它是不可能的亚型病毒使用传统的血清分型方法。分离病毒的基因特征分析结果表明，它是H3 N2亚型，并被命名为A/Turkey/OH/313053/04（H3 N2）。对该病毒8个基因片段的系统发育分析表明，A/Turkey/OH/313053/04（H3 N2）分离株与自1998年以来在美国猪群中传播的三重耐药H3 N2猪病毒[A/Swine/W1/14094/99（H3 N2）]关系最密切，其中包含来自禽、猪和人病毒的基因片段。本研究中从火鸡中分离的A/Turkey/OH/313053/04（H3 N2）被归类为低致病性禽流感A病毒，因为它仅引起产蛋量下降，具有轻微的其他临床体征，并且没有死亡。
Five 34-wk-old turkey breeder layer flocks in separate houses of 2550 birds each in a single farm in Ohio experienced a drop in egg production from late January to early February 2004. Tracheal swabs (n = 60), cloacal swabs (n = 50), and convalescent sera (n = 110) from the flocks were Submitted to the laboratory for diagnostics. Virus isolation was attempted in specific-pathogen free embryonating chicken eggs and Vero and MDCK cells. Virus characterization was performed using agar gel immunodiffusion, the hemagglutination test, the hemagglutination inhibition rest, the virus neutralization test, reverse transcription-polymerase chain reaction, sequencing, and phylogenetic analysis. A presumptive influenza virus was successfully propagated and isolated on the first passage in MDCK cells, but initially not in Vero cells or specific-pathogen free chicken embryos. After two passages in MDCK cells, it was possible to propagate the isolate in specific-pathogen free chicken embryos. Preliminary sequence analysis of the isolated virus confirmed that it was influenza A virus with almost 100% (235/236) identity with the matrix gene of a swine influenza A virus, A/Swine/Illinois/100084/01 (H1N2). However, it was not possible to Subtype the virus using conventional serotyping methods. The results of genetic characterization of the isolated virus showed that it was the H3N2 subtype and was designated as A/Turkey/OH/313053/04 (H3N2). Phylogenetic analysis of the eight gene segments of the virus showed that A/Turkey/OH/313053/04 (H3N2) isolate was most closely related to the triple-reassortant H3N2 swine viruses [A/Swine/W1/14094/99 (H3N2)] that have been circulating among pigs in the United States since 1998, which contains gene segments from avian, swine, and human viruses. The A/Turkey/OH/313053/04 (H3N2) isolated from turkeys in this study was classified as a low pathogenic avian influenza A virus because it only caused a drop in egg production with minor other clinical signs and no mortality.