A Case of Co-infection by Chicken Parvovirus, Infectious Anemia Virus, and Infectious Bronchitis Virus on Broilers
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摘要:
目的 对福建省南平市某肉鸡场不明病因造成肉鸡死亡的病原进行确诊。 方法 通过对鸡传染性支气管炎病毒（Infectious bronchitis virus，IBV）、腺病毒（Fowl adenovirus，FadV）、鸡细小病毒（Chicken parvovirus，ChPV）、新城疫病毒（Newcastle disease virus，NDV）、鸡传染性贫血病毒（Chicken infectious anemia virus，CIAV）、传染性法氏囊病毒（Infectious bursal Disease virus，IBDV）、喉气管炎病毒（Laryngotracheitis virus，LTV）、禽肾炎病毒（Avian nephritis virus，ANV）等病原进行PCR检测，并对扩增片段进行克隆测序，利用生物信息学软件对测序结果进行分析。 结果 PCR检测结果及测序结果显示：鸡细小病毒ChPV、鸡传染性支气管炎病毒IBV、鸡传染性贫血病病毒CIAV为阳性，其余病原检测结果均为阴性。同源性分析结果显示分离株与ChPV的同源性为92.17%～100%，与IBV的同源性为76.9%～84.3%，与CIAV的同源性为91.19%～92.14%；遗传进化树分析结果显示分离到的IBV毒株及CIAV毒株均独处于一个单独的分支，分离到的ChPV毒株与广西参考株亲缘关系较近。 结论 从南平市某鸡场检测到ChPV、IBV、CIAV等3种病原的共感染病例，分离到的IBV毒株的S1基因及CIAV毒株的VP1基因可能已经发生变异。 Abstract:Objective Pathogens causing broiler death by unknown etiology at a chicken farm in Nanping, Fujian were investigated. Methods To establish standard references for this investigation, pathogens on chicken, such as, infectious bronchitis virus (IBV), fowl adenovirus(FadV), chicken parvovirus (ChPV), newcastle disease virus(NDV), chicken infectious anemia virus (CIAV), chicken infectious bursal virus(IBDV), laryngotracheitis virus(LTV), and avian nephritis virus (ANV), were detected by PCR and the amplified fragments cloned, sequenced, and analyzed using bioinformatics software. Results The clinic specimens were found positive on ChPV, IBV, and CIAV but negative on the other viral pathogens. The homology of the isolates was 92.17%–100% with ChPV, 76.9%–84.3% with IBV, and 91.19%–92.14% with CIAV. The isolated IBV and CIAV strains were in their own unique branch, while ChPV closely related to a Guangxi strain as shown by the phylogenetic analysis. Conclusion A case of simultaneous co-infection by ChPV, IBV, and CIAV at a chicken farm was confirmed. The S1 gene of the isolated IBV and the VP1 in the CIAV strain appeared to have mutated prior to the infection incidence. -
图 3 PCR扩增结果
注：M，2 000 Marker；1，阳性对照；2，阴性对照；3，检测样本。
Figure 3. PCR amplification results
Note: M, 2 000 DNA marker; 1, positive control; 2, negative control; 3, clinic specimens.
表 1 所用引物及其序列
Table 1. Primers and sequences applied
病原
Pathogen引物序列（5′→3′）
Primer sequences（5′→3′）目的片段
Target fragment/bpNDV 5′-CAGTGAAGCATGTGAAAGCACC-3′ 365 5′-GAGTACATCACTGAGCCCGACA-3′ IBV 5′-GGCCAGGGTTTTATCACTAA-3′ 215 5′-CGTGAAGTAAGAATGCCAAC-3′ LTV 5′-TTCCGAGATCGAAGAAGTGAG-3′ 567 5′-ACTCTGGTGGCAAGTATCCTGT-3′ IBDV 5′-GACACCCTAGAGAAGCACAC-3′ 509 5′-CTACTGTCACATGTTGCTACC-3′ CIAV 5′-GACTGTAAGATGGCAAGACGAGCTC-3′ 675 5′-GGCTGAAGGATCCCTCATTC-3′ ChPV 5′-GATCCTCAGCGAGTCGAGTG-3′ 937 5′-CCTTGTGCCCCCTGATGATT-3′ ANV 5′-TTCCAATGCCAAAAGTGCAA-3′ 490 5′-AATAGGTGAACCCGATGTCC-3′ 
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表 2 鸡细小病毒、鸡传染性支气管炎病毒、鸡传染性贫血病毒参考株
Table 2. Reference strains of ChPV, IBV, and CIAV
病毒
Virus参考株
Reference
strains登录号
Locus分离地
Isolate site分离时间
Isolate
time病毒
Virus参考株
Reference
strains登录号
Locus分离地
Isolate site分离时间
Isolate
time鸡细小病毒
ChPVChPV 841 KM598417 USA 2015 IS-1494 MG233398 Iran 2019 GX-CH-PV-12 KX133419 Guangxi, China 2017 1939/DMV MK878536 USA 2019 ChPV 528-HUN-2009 GQ281293 Hungary 2016 PR05 MK957244 Brazil 2000 ChPV 736 KM598415 USA 2015 TW I MN128087 Taiwan, China 1998 ChPV ABU-84 EU304808 Hungary 2008 TW II MN128088 Taiwan, China 1995 ChPV ABU-P1 GU214704 Hungary 2010 Italy02 MN548288 UK 2011 ChPV ADL120035 KJ486490 South Korea 2015 QX MN548289 UK 2011 ChPV ADL120686 KJ486491 South Korea 2015 GA08（GI-27） MN599049 USA 2015 CH-CH-PV-2 KX084400 Guangxi, China 2017 1639（GI-17） MN757859 Costa Rica 2016 CH-CH-PV-4 KX084401 Guangxi, China 2017 H52 vaccine MP656388 Germany 2020 GX-CH-PV-6 KX133427 Guangxi, China 2017 4/91 vaccine MP657738 Germany 2020 GX-CH-PV-7 KU523900 Guangxi, China 2017 GI-19 KC577395 Guangdong, China 1993 GX-CH-PV-8 KX133415 Guangxi, China 2017 GI-21 DQ064806 Spain 1997 GX-CH-PV-9 KX133416 Guangxi, China 2017 鸡传染性
贫血病毒
CIAVGX1905 MN103404 Guangxi, China 2019 GX-CH-PV-11 KX133418 Guangxi, China 2017 SD1505 KU645523 Shandong, China 2015 GX-CH-PV-14 KX133421 Guangxi, China 2017 JS1503 KU645509 Jiangsu, China 2015 GX-CH-PV-15 KX133422 Guangxi, China 2017 SD1502 KU645508 Shandong, China 2015 GX-CH-PV-16 KX133423 Guangxi, China 2017 CAV-SD189106 MT332128 Shandong, China 2015 GX-CH-PV-17 KX133424 Guangxi, China 2017 CAV-JL186101 MT332122 Jilin, China 2018 GX-CH-PV-18 KX133425 Guangxi, China 2017 CAV-HuN169602 MT332117 Hunan, China 2016 GX-CH-PV-19 MG602509 Guangxi, China 2017 CAV-HuN169002 MT332116 Hunan, China 2016 GX-CH-PV-20 MG602510 Guangxi, China 2017 CAV-HLJ1611605 MT332114 Heilongjiang, China 2016 GX-CH-PV-21 MG602511 Guangxi, China 2017 CAV-BJ171403 MT332104 Beijing, China 2017 GX-CH-PV-22 MG602512 Guangxi, China 2017 1011PT02 KY888891 Taiwan, China 2010 GX-CH-PV-23 MG602513 Guangxi, China 2017 GX1904P MN103403 Guangxi, China 2019 GX-CH-PV-24 MG602514 Guangxi, China 2017 1009PT01 KY888890 Taiwan, China 2010 GX-CH-PV-25 MG602515 Guangxi, China 2017 HLJ15166 KY486142 Heilongjiang, China 2015 GX-CH-PV-26 MG602516 Guangxi, China 2017 HLJ15165 KY486141 Heilongjiang, China 2015 GX-CH-PV-27 MG602517 Guangxi, China 2017 HLJ14101 KY486136 Heilongjiang, China 2014 GX-CH-PV-28 MG602518 Guangxi, China 2017 BS-C2 KX447634 Shandong, China 2017 GX-CH-PV-29 MG602519 Guangxi, China 2017 NGP-7 KX377127 India 2012 GX-CH-PV-30 MG602520 Guangxi, China 2017 HN1405 KU645520 Henan, China 2014 USP 362-3 MK440128 Brazil 2019 SD1512 KU645506 Shandong, China 2015 火鸡细小
病毒
TuPVTuPV 1078 GU214705 USA 2010 CAVNAMKS1 KP120758 India 2009 GX-Tu-PV-2 KX084397 Guangxi, China 2017 EB7K MT259316 Turkey 2016 GX-Tu-PV-3 KX084398 Guangxi, China 2017 CAV-HLJ187007 MT332110 Heilongjiang, China 2018 鸡传染性
支气管炎
病毒
IBVArkDPI EU418975 USA 2009 CAV-JL184202 MT332120 Jilin, China 2018 H52 EU817497 Zhejiang, China 2009 CAV-JL184203 MT332121 Jilin, China 2018 Mass41 FJ904713 USA 2006 CAV-JL186105 MT332123 Jilin, China 2018 Cal 1995 FJ904714 USA 2011 CAV-SD194605 MT332129 Shandong, China 2019 Cal56b GU393333 USA 2011 CAV-SD194607 MT332131 Shandong, China 2019 H120 GU393335 USA 2011 CAV-SD194608 MT332132 Shandong, China 2019 Mass KJ435283 Heilongjiang, China 2016 N7 MK887165 Guangdong, China 2016 973B KP118889 Heilongjiang, China 2015 N6 MK887166 Guangdong, China 2016 LDT3-A KR608272 Heilongjiang, China 2016 N4 MK887168 Guangdong, China 2016 LDT3 KT852992 Heilongjiang, China 2016 N2 MK887170 Guangdong, China 2016 LH1 KX185059 Heilongjiang, China 2017 1510TW MN299309 Taiwan, China 2015 LX4 KX302866 Heilongjiang, China 2017 1637TW MN299310 Taiwan, China 2016 GI-23 KY805846 Egypt 2017 1725PT MN299311 Taiwan, China 2016
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