2013年臺灣地區豬流行性感冒病毒分離及親緣關係分析

Virus Isolation and Phylogenetic Analysis for Swine Influenza Virus in Taiwan in 2013

張致中(Chih-Chung Chang)

指導教授 ： 邱明堂 ； 林昭男

國立屏東科技大學/農學院/獸醫學系所/碩士(2014年)

https://doi.org/10.6346/NPUST.2014.00315

摘要

豬流行性感冒病毒 ( swine influenza virus; SIV ) 屬於A型流行性感冒病毒屬 ( genus Influenzavirus A )，正黏液病毒科 ( family Orthomyxoviridae )。A型流行性感冒病毒 ( Influenza A virus; IAV ) 於人類、鳥類及哺乳動物為一具高度傳染性之病原。根據病毒表面之血球凝集素 ( hemagglutinin; HA ) 及神經胺酸酶 ( neuraminidase; NA ) 抗原性之不同區分出17種HA亞型 ( subtype ) 及9種NA亞型。而豬隻被認為在IAV扮演一個混合容器 ( mix vessel ) 的角色，因為家禽流行性感冒病毒及人類流行性感冒病毒皆可於豬隻體內複製。為了進行臺灣SIV之序列分析，於2013年1月至2014年1月共自10個豬場採集219個鼻腔拭子樣本及87個豬場收集183個肺臟組織樣本進行SIV的分離及PCR之檢測。結果顯示，以PCR檢測鼻腔拭子樣本及肺臟組織樣本之陽性率分別為5.5% ( 12/219 ) 及47% ( 86/183 )。而SIV自鼻腔拭子樣本及肺臟組織樣本之分離率分別為5.9% ( 13/219 ) 及3.3% ( 6/183 )。選取14株SIV病毒株進行HA、NA及基質 ( matrix; M ) 基因之定序及序列分析。此14株SIV病毒株分別為3株H1N1病毒株、5株H1N2病毒株、5株H3N1病毒株及1株H3N2 病毒株。8株H1亞型SIV病毒株分別劃分於典型豬流行性感冒病毒 ( classical SIV )( 82.7-88.6% ) 和2009年泛流行性感冒病毒 ( pandemic 2009 IV )( 95.3-97.6% ) 之演化分支 ( clade ) 內；6株H3亞型SIV病毒株獨自形成一個新的演化分支，並與早期人類流行性感冒病毒 ( early human IV )( 85.1-85.5% ) 最為相似。8株N1亞型SIV病毒株被劃分於classical SIV ( 84.3-89.5% ) 之演化分支內；6株N2亞型SIV病毒株獨自形成一個新的演化分支，並與early human IV ( 88.3-89.1% ) 最為相似。14株SIV病毒株的M基因序列皆被劃分於pandemic 2009 IV ( 97.8-98.8% )。綜上所述，臺灣豬群內可見H1N1、H1N2、H3N1及H3N2等4種亞型之SIV同時存在，甚至於同一來源豬場可分離到超過1種亞型之SIV。分析本研究所分離之SIV病毒株，發現其HA、NA及M基因出現由classical SIV、pandemic 2009 IV及early human IV重排之現象。

關鍵字

豬流行性感冒 ； 病毒分離 ； 全長序列分析 ； 臺灣

並列摘要

Swine influenza virus (SIV) belongs to genus Influenzavirus A of the family Orthomyxoviridae. Influenza A virus (IAV) is a highly infectious respiratory pathogen in humans, birds and mammals. Based on the differences in antigenicity of hemagglutinin (HA) and neuraminidase (NA), it can be subdivided into 17 HA subtypes and 9 NA subtypes. The pigs have been considered as a “mixing vessel” for IAV, because it supports replication of avian and human influenza viruses. In order to analysis SIV sequences of pigs in Taiwan, SIVs were try to isolated and detected by PCR from 219 nasal swabs from 10 pig farms and 183 lung tissues from 87 pig farms from January 2013 to January 2014. The results show the PCR positive rate of nasal swabs and lung tissues were 5.5% (12/219) and 47.0% (86/183), respectively. SIV isolation rate of nasal swabs and lung tissues were 5.9% (13/219) and 3.3% (6/183), respectively. Fourteen selected SIV isolates were subsequently sequenced and analyzed the HA, NA and M gene. The fourteen isolated SIVs belonged to the subtypes H1N1 (n=3), H1N2 (n=5), H3N1 (n=5) and H3N2 (n=1). The sequences of eight H1 subtype isolates were clustered with classical SIVs (82.7-88.6%) and pandemic 2009 influenza viruses (IVs) (95.3-97.6%), respectively. The sequences of six H3 isolates were formed a new clade and were most similar to early human IV (85.1-85.5%). The sequences of eight N1 subtype isolates were clustered with classical SIVs (84.3-89.5%); the sequences of six N2 subtype sequences were formed a new clade and were most similar to early human IV (88.3-89.1%). The M gene sequences of all isolates were similar to pandemic 2009 IVs (97.8-98.8%). In conclusion, there were four subtypes (H1N1, H1N2, H3N1 and H3N2) SIVs co-circulation in pigs in Taiwan, even more than one subtype SIVs isolated from the same pig farm. HA, NA and M genes of the SIV isolates were reasserted with classical SIV, pandemic 2009 IV and early human IV.

並列關鍵字

swine influenza virus ； virus isolation ； full-lenth sequence analysis ； Taiwan

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