内蒙古绵羊梅迪-维斯纳病的病理组织学和巢式PCR诊断
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摘要
为检测内蒙古地区是否存在梅迪-维斯纳病(Maedi-Visna dieas,eMVD),本试验以内蒙古呼浩特市周边某屠宰场采集的12份绵羊病肺组织为研究对象,采用组织病理学、巢式PCR和测序分析等方法进行检测。组织病理学结果显示:肺脏发生纤维化,并且肺泡腔内Ⅱ型肺泡上皮增生以及间质增宽,支气管和细支气管周围可见较多的淋巴滤泡增生;巢式PCR结果显示:病肺组织中有梅迪-维斯纳病毒(Maedi-Visna viru,sMVV)G ag基因序列的特异性扩增,且与小反刍兽疫、绵羊肺腺瘤病毒以及支原体均无交叉反应。以上结果表明,内蒙古存在MVD。
In order to detect the presence of Maedi-Visna Diease(MVD) in Hohhot,Inner Mongolia,this study used 12 sheep lung tissues collected from a slaughterhouse around Hohhot,to study histopathology Nested PCR and sequencing analysis were performed.Histopathological results showed that 3 diseased lung tissues had characteristic pathological changes of MVD,fibrosis of the lungs,alveolar epithelial hyperplasia and interstitial hyperplasia in the alveolar cavity,and more lymphoid follicles around the bronchi and bronchioles.The gag gene sequence of Maedi-Visna Virus(MVV) was specificly amplified by the nested PCR from the 3 diseased lung tissues,but not from the ovine samples with lung adenovirus infection,peste des petits ruminants virus infection or Mycoplasma infection,respectively.The abovementioned results indicated that MVD exists in Inner Mongolia,China.
In order to detect the presence of Maedi-Visna Diease(MVD) in Hohhot,Inner Mongolia,this study used 12 sheep lung tissues collected from a slaughterhouse around Hohhot,to study histopathology Nested PCR and sequencing analysis were performed.Histopathological results showed that 3 diseased lung tissues had characteristic pathological changes of MVD,fibrosis of the lungs,alveolar epithelial hyperplasia and interstitial hyperplasia in the alveolar cavity,and more lymphoid follicles around the bronchi and bronchioles.The gag gene sequence of Maedi-Visna Virus(MVV) was specificly amplified by the nested PCR from the 3 diseased lung tissues,but not from the ovine samples with lung adenovirus infection,peste des petits ruminants virus infection or Mycoplasma infection,respectively.The abovementioned results indicated that MVD exists in Inner Mongolia,China.
引文
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[10]丁忠庆.绵羊梅迪-维斯纳病毒核酸疫苗的制备及免疫学研究[D].哈尔滨:东北农业大学,2006.DING Zhong-qing.Preparation of Medici-visna virus DNA vaccine and its immunological research[D].Harbin:Northeast Agricultural University,2006.(in Chinese)
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[13]徐军,孙志华,刘娟,等.梅迪-维斯纳病毒和羊痘病毒多联实时定量PCR检测方法的建立及初步应用[J].石河子大学学报(自然科学版),2012(04):448-451.XU Jun,SUN Zhi-hua,LIU Juan,et al.Establishment of a multiplex real-time flourescence quantitative PCR assay for detection of Brucella and Mycobacterium tuberculosis[J].Journal of Shihezi University:Natural Science,2012(04):448-451.(in Chinese)
[14]张太翔,凌宗帅,孙涛,等.梅迪-维斯纳病毒实时荧光定量PCR检测方法的建立[J].中国预防兽医学报,2011(02):163-165.ZHANG Tai-xiang,LIN Zong-shuai,SUN Tao,et al.Realtime quantitative RT-PCR assay for detection of Maedi-visna virus[J].Chinese Journal of Preveentive Veterinary Medicine,2011(02):163-165.(in Chinese)
[15]EXTRAMIANA A B,GONZALEZ L,CORTABARR A N,et al.E-valuation of a PCR technique for the detection of Maedi-Visna proviral DNA in blood,milk and tissue samples of naturally infected sheep[J].Small Ruminant Research,2002,44(2):109-118.
[16]AZIZI S,TAJBAKHSH E,FATHI F,et al.Maedi in slaughtered sheep:A pathology and polymerase chain reaction study in southwestern Iran[J].Tropical Animal Health and Production,2012,44(1):113-118.
[17]BARQUERO N,DOMENECH A,ARJONA A,et al.Comparison of two PCR and one ELISA techniques for the detection of small ruminant lentiviruses(SRLVs)in milk of sheep and goats[J].Research in Veterinary Science,2013,94(3):817-819.
[18]郭文庆,孙晓琳,冷冉,等.绵羊肺腺瘤病毒巢式RT-PCR检测方法的建立与验证[J].中国兽医科学,2017,47(01):23-30.GUO Wen-qing,SUN Xiao-ling,LENG Ran,et al.Establishment and verification of a nested RT-PCR method for detection of Jaagsiekte sheep retrovirus[J].Chinese Veterinary Science,2017,47(01):23-30.(in Chinese)
[19]孙雨,杨林,王传彬,等.我国11个省份山羊关节炎-脑炎与绵羊进行性肺炎的血清抗体检测[J].中国兽医科学,2018,48(01):34-38.SUN Yu,YANG Lin,WANG Chuan-shan,et al.Serological investigation of caprine arthritis encephalitis and ovine progressive pneumoniina in 11 provinces of China[J].Chinese Veterinary Science,2018,48(01):34-38.(in Chinese)
[2]VIGNER,BARBAN V,QUERAT G,et al.Transcription of visnavi-rus during its lytic cycle:evidence for as equential early and late gene expression[J].Virology,1987,7(161):218-227.
[3]于立新,敖双捷,幺宏强,等.自然感染绵羊肺腺瘤病和绵羊进行性肺炎病例肺脏的病理组织学观察[J].畜牧与饲料科学,2011(06):1-2.YU Li-xin,AO Shuang-jie,YAO Hong-qiang,et al.Histopathological observation of ovine pulmonary adenomatosis and ovine progressive pneumonia in lung[J].Feed and Animal Husbandry,2011(06):1-2.(in Chinese)
[4]邓普辉,简子健,阿合买提,等.绵羊慢病毒北疆株自然感染新疆美利奴羊的病理学[J].中国兽医杂志,2001,37(11):9-11.DENG Pu-hui,JIAN Zi-jian,Ahe Mai-ti,et al.Pathology of Xinjiang Merino sheep naturally infected by sheep lentivirus[J].Chinese Journal of Veterinary Medicine,2001,37(11):9-11.(in Chinese)
[5]EAST N E,ROWE J D,MADEWELL B R,et al.Serologic prevalence of caprine arthritis-encephalitisvirus in California goat dairies[J].Journal of the American Veterinary Medical Association,1987,190(2):182-186.
[6]GUFLER H,MORONI P,CASU S,et al.Seroprevalence,clinical incidence,and molecular and epidemiolo-gical characterisation of small ruminant lenti-virus in the indigenous Passirian goat in northernItaly[J].Archives Virology,2008(8):1581-1585.
[7]GUFLER H,GASTEINER J,LOMBARDO D.Serological study of small ruminant lentivirus in goats in Italy[J]Small Ruminant Research,2007,73(3):169-173.
[8]GLARIA I,REINA R,CRESPO H,et al.Phylogenetic analysis of SRLV sequences from an arthritic sheep outbreak demonstrates the introductionof CAEV-like viruses among Spanish sheep[J].Veterinary Microbiology,2009,138(1-2):156-162.
[9]司俊强.新疆部分地区羊场绵羊肺腺瘤病和梅迪-维斯纳病调查及病理学鉴别诊断[D].石河子:石河子大学,2017.SI Jun-qiang.Investigation and pathological diagnosis of sheep pulmonary adenomatosisand MaediVisna disease in part areas of Xinjiang[D].Shihezi:Shihezi University,2017.(in Chinese)
[10]丁忠庆.绵羊梅迪-维斯纳病毒核酸疫苗的制备及免疫学研究[D].哈尔滨:东北农业大学,2006.DING Zhong-qing.Preparation of Medici-visna virus DNA vaccine and its immunological research[D].Harbin:Northeast Agricultural University,2006.(in Chinese)
[11]HEINRICHS R,WILKINS W,SCHROEDERr G,et al.Prevalence of Maedi-visna in Saskatchewan sheep[J].Canadian Veterinary Journal,2017,58(2):183-186.
[12]BENAVIDES J,FUERTES M,GARCIA-PARIENTE C,et al.Natural cases of visna in sheep with myelitis as the sole lesion in the central nervous system[J].Journal of Comparative Pathology,2006,134(2-3):219-230.
[13]徐军,孙志华,刘娟,等.梅迪-维斯纳病毒和羊痘病毒多联实时定量PCR检测方法的建立及初步应用[J].石河子大学学报(自然科学版),2012(04):448-451.XU Jun,SUN Zhi-hua,LIU Juan,et al.Establishment of a multiplex real-time flourescence quantitative PCR assay for detection of Brucella and Mycobacterium tuberculosis[J].Journal of Shihezi University:Natural Science,2012(04):448-451.(in Chinese)
[14]张太翔,凌宗帅,孙涛,等.梅迪-维斯纳病毒实时荧光定量PCR检测方法的建立[J].中国预防兽医学报,2011(02):163-165.ZHANG Tai-xiang,LIN Zong-shuai,SUN Tao,et al.Realtime quantitative RT-PCR assay for detection of Maedi-visna virus[J].Chinese Journal of Preveentive Veterinary Medicine,2011(02):163-165.(in Chinese)
[15]EXTRAMIANA A B,GONZALEZ L,CORTABARR A N,et al.E-valuation of a PCR technique for the detection of Maedi-Visna proviral DNA in blood,milk and tissue samples of naturally infected sheep[J].Small Ruminant Research,2002,44(2):109-118.
[16]AZIZI S,TAJBAKHSH E,FATHI F,et al.Maedi in slaughtered sheep:A pathology and polymerase chain reaction study in southwestern Iran[J].Tropical Animal Health and Production,2012,44(1):113-118.
[17]BARQUERO N,DOMENECH A,ARJONA A,et al.Comparison of two PCR and one ELISA techniques for the detection of small ruminant lentiviruses(SRLVs)in milk of sheep and goats[J].Research in Veterinary Science,2013,94(3):817-819.
[18]郭文庆,孙晓琳,冷冉,等.绵羊肺腺瘤病毒巢式RT-PCR检测方法的建立与验证[J].中国兽医科学,2017,47(01):23-30.GUO Wen-qing,SUN Xiao-ling,LENG Ran,et al.Establishment and verification of a nested RT-PCR method for detection of Jaagsiekte sheep retrovirus[J].Chinese Veterinary Science,2017,47(01):23-30.(in Chinese)
[19]孙雨,杨林,王传彬,等.我国11个省份山羊关节炎-脑炎与绵羊进行性肺炎的血清抗体检测[J].中国兽医科学,2018,48(01):34-38.SUN Yu,YANG Lin,WANG Chuan-shan,et al.Serological investigation of caprine arthritis encephalitis and ovine progressive pneumoniina in 11 provinces of China[J].Chinese Veterinary Science,2018,48(01):34-38.(in Chinese)