番鸭中网状内皮组织增生症病毒感染的检测

万春和; 刘荣昌; 程龙飞; 傅光华; 施少华; 陈红梅; 傅秋玲; 黄瑜

doi:10.19303/j.issn.1008-0384.2016.10.001

番鸭中网状内皮组织增生症病毒感染的检测

doi: 10.19303/j.issn.1008-0384.2016.10.001

福建省农业科学院畜牧兽医研究所/福建省畜禽疫病防治工程技术研究中心/福建省禽病防治重点实验室, 福建福州 350013

基金项目:

国家水禽产业技术体系项目 CARS-43

福建省畜禽疫病防控技术重大研发平台 2014N2003

福建省优良蛋鸭品种与设施化养殖创新产业化工程项目 2014-2016

福建省农业科学院畜牧兽医研究所基金 MYQJ2015（S）-3

详细信息

作者简介:
万春和(1982-), 男, 博士, 助理研究员, 主要从事水禽病毒分子生物学研究(E-mail:chunhewan@126.com)

通讯作者:
黄瑜(1965-), 男, 博士, 研究员, 主要从事畜禽传染病防控研究(E-mail:huangyu_815@163.com)

中图分类号: S852
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出版历程
- 收稿日期: 2016-08-03
- 修回日期: 2016-09-17
- 刊出日期: 2016-10-28

Evidence of Viral Reticuloendotheliosis in Muscovy Ducks

Institute of Animal Husbandry and Veterinary Medicine of Fujian Academy of Agricultural Sciences/Fujian Animal Diseases Control Technology Development Center/Fujian Provincial Key Laboratory for Avian Diseases Control and Prevention, Fuzhou, Fujian 350013, China

摘要

摘要: 网状内皮组织增生症病毒（reticuloendotheliosis virus，REV）是一种可感染多品种禽的免疫抑制病病原，禽类感染REV后会导致机体发生免疫抑制并由此引发多种病原的继发感染（或混合感染），给养禽业造成极大损失。本研究利用针对REV基因组长末端重复序列（long terminal repeat，LTR）的特异性引物，对21份2013年收集的福建（12份）和浙江（9份）两省的番鸭源病料（肝脏和脾脏组织）进行检测。其中来源于福建漳州（1株，记为REV-FJ-MD01）和浙江宁波（1株，记为REV-ZJ-MD01）的番鸭源病料检测到REV感染阳性。将目的片段经克隆后进行序列测定分析发现，REV-FJ-MD01和REV-ZJ-MD01在该扩增区域的核苷酸同源性为100%，与GenBank中的REV参考株（除鸽源REV和鸭源713株仅1个碱基差异外）核苷酸同源性也为100%；从遗传进化关系可以看出，REV-FJ-MD01和REV-ZJ-MD01与REV参考株遗传进化较近，处于相同的遗传进化分支。本研究首次证实在福建番鸭中存在REV感染。
- 网状内皮组织增生症病毒 /
- 番鸭 /
- 感染
Abstract: The diseases caused by reticuloendotheliosis virus (REV) are generally characterized by immunosuppression occurred to avian species. They can result in the increased susceptibility to secondary bacterial or viral infections and poor immune responses to vaccines on the infected birds. An epidemic of the viral diseases can lead to disastrous impacts for the poultry industry. This study employed the specific primers-pair targeting the long terminal repeat of the virus genomic sequence for an investigation on 21 samples from the infected Muscovy ducks (12 from Fujian and 9 from Zhejiang) collected in 2013. Among the samples, one from Fujian (designated as strain REV-FJ-MD01) and another from Zhejiang (designated as strain REV-ZJ-MD01) were identified for further study. The positive fragments were harvested, cloned, and sequenced. The obtained sequence shared a perfect nucleotide identity with other GenBank REV isolates, except the pigeon-origin REV and the duck-origin REV strain 173, which differed on only one nucleotide. The genetic evolutionary relationship between the two strains and REV appeared close. Therefore, it was concluded that REV infection existed in the Muscovy ducks in Fujian.
- reticuloendotheliosis virus /
- muscovy duck /
- infection

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图 1 临床样品的PCR扩增

注：1~3为REV-FJ-MD01病料MDV、FPV和REV检测；4~6为REV-ZJ-MD01病料MDV、FPV和REV检测；7为阴性对照；M为DL 2 000 Marker。

Figure 1. PCR detection on clinic samples

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图 2 LTR序列遗传进化分析

Figure 2. Phylogenetic analysis based on LTR sequences

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表 1 本研究使用的引物

Table 1. Primers employed

病原	引物名称	引物序列	针对基因	大小/bp	文献
REV	REV-F1	GCCTTAGCCGCC ATTGTA	LTR	383	^[11]
	REV-R1	CCAGCCTACACC ACGAACA
MDV	MDV-F1	GGATCGCCCACC ACGATTACTACC	Meq	400	^[12]
	MDV-R1	ACTGCCTCACAC AACCTCATCTCC
FPV	FPV-F1	TCAGCAGGTGCT AAACAACA	P4b	579	^[13]
	FPV-R1	CGGTAGCTTAAC GCCGAATA

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表 2 参考株信息

Table 2. Information on reference strains

GenBank号	名称	宿主	时间	地区	文献
FJ439119	3410/06	goose	2006	台湾	^[4]
FJ439120	3337/05	chicken	2005	台湾	^[4]
GQ415646	HLJR0901	chicken	2009	黑龙江	^[14]
JQ804915	1105	Muscovy duck	2011	浙江	^[10]
KJ756349	104865	turkey	2014	USA	不详
KU204703	GDBL1402	pigeon	2014	广东	^[15]
KU204702	GDBL1401	pigeon	2014	广东	^[15]
M22223	713	duck	1973	USA	不详
KT833852^a	rMd5	none	2016	USA	不详
注：表示MDV整合有REV的LTR区序列。

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参考文献(21)

[1]	高玉龙, 秦立廷, 王笑梅.家禽病毒性免疫抑制病流行特点与防控对策[J].中国家禽, 2012, 34(15):5-11. http://www.cnki.com.cn/Article/CJFDTOTAL-ZGJQ201215003.htm
[2]	张颖, 刁有祥, 宋晓娜, 等.商品肉鸭免疫抑制病病原体共感染情况调查[J].中国兽医学报, 2013, 33(6):828-832. http://www.cnki.com.cn/Article/CJFDTOTAL-ZSYX201306005.htm
[3]	陈翠腾, 傅光华, 黄瑜.家禽病毒性免疫抑制病研究进展[J].畜牧与兽医, 2014, 46(7):126-129. http://www.cnki.com.cn/Article/CJFDTOTAL-XMYS201407037.htm
[4]	LIN C, CHEN C, WANG C, et al. Isolation, identification, and complete genome sequence of an avian reticuloendotheliosis virus isolated from geese[J]. Vet Microbiol, 2009, 136(3-4):246-249. doi: 10.1016/j.vetmic.2008.11.017
[5]	邓小芸, 祁小乐, 高玉龙, 等.禽网状内皮组织增生症流行现状及检测技术研究进展[J].动物医学进展, 2010, 31(10):93-96. http://www.cnki.com.cn/Article/CJFDTOTAL-DYJZ201009023.htm
[6]	PURCHASE H G, WITTER R L. The reticuloendotheliosis viruses[J]. Curr Top Microbiol Immunol, 1975, 71:103-124. https://www.ncbi.nlm.nih.gov/pubmed/53119
[7]	何宏虎, 陈溥言, 蔡宝祥.禽网状内皮组织增殖病病毒的分离鉴定[J].中国畜禽传染病, 1988, (2):1-2. http://www.cnki.com.cn/Article/CJFDTOTAL-ZGXQ198802000.htm
[8]	姜莉莉.东北地区野生鸟类主要禽免疫抑制病病原感染情况调查及与REV PR相互作用宿主蛋白的筛选[D].哈尔滨:东北林业大学, 2014. http://cdmd.cnki.com.cn/Article/CDMD-10225-1014411653.htm
[9]	倪楠, 崔治中.鸭群中REV感染的流行病学调查[J].微生物学报, 2008, 48(4):514-519. http://www.cnki.com.cn/Article/CJFDTOTAL-WSXB200804022.htm
[10]	JIANG T, LU X, YUAN Y, et al. Complete genomic sequence of a Muscovy duck-origin reticuloendotheliosis virus from China[J]. J Virol, 2012, 86(23):13140-13141. doi: 10.1128/JVI.02531-12
[11]	JIANG L, QI X, GAO Y, et al. Molecular characterization and phylogenetic analysis of the reticuloendotheliosis virus isolated from wild birds in Northeast China[J]. Vet Microbiol, 2013, 166(1-2):68-75. doi: 10.1016/j.vetmic.2013.05.008
[12]	RAJA A, DHINAKAR RAG G, BHUVANESWARI P, et al. Detection of virulent Marek's disease virus in poultry in India[J]. Acta Virol, 2009, 53(4):255-260. doi: 10.4149/av_2009_04_255
[13]	HUW LEE L, HWA LEE K. Application of the polymerase chain reaction for the diagnosis of fowl poxvirus infection[J]. J Virol Methods, 1997, 63(1-2):113-119. doi: 10.1016/S0166-0934(96)02119-2
[14]	DENG X, QI X, WU G, et al. Construction and characterization of the infectious clone of Reticuloendotheliosis virus carrying a genetic marker[J]. Virus Res, 2012, 167(2):146-151. doi: 10.1016/j.virusres.2012.02.011
[15]	ZHAI S, CHEN S, LIN T, et al. Emergence of reticuloendotheliosis virus in pigeons in Guangdong Province, Southern China[J]. Arch Virol, 2016, 161(7):2007-2011. doi: 10.1007/s00705-016-2870-3
[16]	MOTHA MX. Clinical effects, virological and serological responses in chickens following in-ovo inoculation of reticuloendotheliosis virus[J]. Vet Microbiol, 1987, 14(4):411-417. doi: 10.1016/0378-1135(87)90031-9
[17]	JIANG L, DENG X, GAO Y, et al. First isolation of reticuloendotheliosis virus from mallards in China[J]. Arch Virol, 2014, 159(8):2051-2057. doi: 10.1007/s00705-013-1821-5
[18]	LI J, DONG X, YANG C, et al. Isolation, identification, and whole genome sequencing of reticuloendotheliosis virus from a vaccine against Marek's disease[J]. Poult Sci, 2015, 94(4):643-649. doi: 10.3382/ps/pev034
[19]	KOO B S, LEE H R, JEON E O, et al. An outbreak of lymphomas in a layer chicken flock previously infected with fowlpox virus containing integrated reticuloendotheliosis virus[J]. Avian Dis, 2013, 57(4):812-817. doi: 10.1637/10551-041113-Case.R1
[20]	崔治中.禽反转录病毒与DNA病毒间的基因重组及其流行病学意义[J].病毒学报, 2006, 22(2):150-154. http://www.cnki.com.cn/Article/CJFDTOTAL-BDXB200602014.htm
[21]	黄瑜, 祁保民, 彭春香, 等.鸭的免疫抑制病[J].中国兽医杂志, 2010, 46(7):48-50. http://www.cnki.com.cn/Article/CJFDTOTAL-ZSYZ201007024.htm