荧光定量RT-PCR方法检测HP-PRRSV JXA1分离株在藏猪、藏梅猪和大约克夏猪体内的动态分布
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摘要
为了探明高致病性猪繁殖与呼吸综合征病毒(HP-PRRSV)JXA1分离株感染藏猪、藏梅猪和大约克夏猪后,病毒在猪体内的分布规律,本研究利用荧光定量RT-PCR技术检测3个品种猪感染4×105TCID50/m L的JXA1分离株后0、4、7和14 d体内的15个器官(心、肝、脾、肺、肾、大脑、小肠、颌下腺、颌下淋巴结、甲状腺、胸腺、支气管淋巴结、肠系膜淋巴结、腹股沟淋巴结和扁桃体)的病毒载量。结果表明,在整个感染期间,藏梅猪和大约克夏猪的15个器官中均检测到JXA1分离株,而藏猪的心、肝、脾、肾、小肠、颌下腺和甲状腺以及感染后14 d的脑中未检测到JXA1分离株;在JXA1分离株感染3个品种猪4、7和14 d,藏猪病毒载量最多的器官分别是扁桃体、腹股沟淋巴结和颌下淋巴结,藏梅猪分别是脾、脾和肺,大约克夏猪分别为胸腺、肺和颌下淋巴结。本研究证实了JXA1分离株在3个品种猪体内的分布规律和器官嗜性均存在明显差异。
In order to investigate viral distribution in PRRSV-infected Tibetan,Zang Mei black and Yorkshire pigs,we used 1 m L 4×105 TCID50/ml of HP-PRRSV isolate to infect Tibetan,Zang Mei black and Yorkshire pigs( 15 for each breed) and measured viral RNA loads in each of the fifteen tested organs of the pigs on 0,4,7,14 days after PRRSV challenge. The 15 tested organs were heart,liver,spleen,lung,kidney,brain,small intestine,mesenteric lymph node,groin lymph node,submandibular gland,mandibular lymph node,thyroid,thymus,bronchial lymph nodes and tonsil. The results were that viral RNA was identified in all the fifteen organs of Zang Mei black and Yorkshire pigs,but not in the heart,liver,spleen,kidney,small intestine,submandibular gland and thyroid of the Tibetan pigs during infection.The maximum viral RNA loads in the organs of the three pig breeds 4,7,and 14 days after infection were evidently different,with the highest loads in the amygdala,inguinal lymph nodes and lymph nodes of lower jaw of the Tibetan pigs,in the spleen and lung of Zang Mei black pigs,and in the thymus,lung and lower jaw lymph of the Yorkshire pigs. To sum up,HP-PRRSV infection results in different distribution patterns of virus in the three pig breeds as a whole and in different organs of individual breeds of the pigs.
In order to investigate viral distribution in PRRSV-infected Tibetan,Zang Mei black and Yorkshire pigs,we used 1 m L 4×105 TCID50/ml of HP-PRRSV isolate to infect Tibetan,Zang Mei black and Yorkshire pigs( 15 for each breed) and measured viral RNA loads in each of the fifteen tested organs of the pigs on 0,4,7,14 days after PRRSV challenge. The 15 tested organs were heart,liver,spleen,lung,kidney,brain,small intestine,mesenteric lymph node,groin lymph node,submandibular gland,mandibular lymph node,thyroid,thymus,bronchial lymph nodes and tonsil. The results were that viral RNA was identified in all the fifteen organs of Zang Mei black and Yorkshire pigs,but not in the heart,liver,spleen,kidney,small intestine,submandibular gland and thyroid of the Tibetan pigs during infection.The maximum viral RNA loads in the organs of the three pig breeds 4,7,and 14 days after infection were evidently different,with the highest loads in the amygdala,inguinal lymph nodes and lymph nodes of lower jaw of the Tibetan pigs,in the spleen and lung of Zang Mei black pigs,and in the thymus,lung and lower jaw lymph of the Yorkshire pigs. To sum up,HP-PRRSV infection results in different distribution patterns of virus in the three pig breeds as a whole and in different organs of individual breeds of the pigs.
引文
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[2]Metwally S,Mohamed F,Faaberg K,et al.Pathogenicity and molecular characterization of emerging porcine reproductive and respiratory syndrome virus in Vietnam in 2007[J].Transboundary and Emerging Diseases,2010,57(5):315-329.
[3]Rossow KD,Benfield DA,Goyal SM,et al.Chronological immunohistochemical detection and localization of porcine reproductive and respiratory syndrome virus in gnotobiotic pigs[J].Veterinary Pathology,1996,33(5):551-556.
[4]Li L M,Zhao Q,Ge X N,et al.Chinese highly pathogenic porcine reproductive and respiratory syndrome virus exhibits more extensive tissue tropism for pigs[J].Virology Journal,2012,9(1):1-6.
[5]Sur JH,Doster AR,Christian JS,et al.Porcine reproductive and respiratory syndrome virus replicates in testicular germ cells,alters spermatogenesis,and induces germ cell death by apoptosis[J].Journal of Virology,1997,71(12):9170-9179.
[6]Han K,Seo HW,Oh Y,et al.Comparison of the virulence of European and North American genotypes of porcine reproductive and respiratory syndrome virus in experimentally infected pigs[J].Veterinary Journal,2013.195(3):313-318.
[7]Do TD,Park C,Choi K,et al.Comparison of pathogenicity of highly pathogenic porcine reproductive and respiratory syndrome virus between wild and domestic pigs[J].Veterinary Research Communications,2015,39(1):79-85.
[8]Kang R M,Ji G S,Yang X,et al.Investigation on host susceptibility of Tibetan pig to infection of porcine reproductive and respiratory syndrome virus through viral challenge study[J].Veterinary Microbiology.2016,183(5):62-68.
[9]Zhou Y H,Yang X,Wang H N,et al.Molecular characterization of a complete genome and 12 Nsp2 genes of PRRSVof southwestern China[J].Food and Environmental Virology,2012,4(3):102-114.
[10]Muhammad Jasim Uddin,Mehmet Ulas Cinar,Dawit Tesfaye,et al.Age-related changes in relative expression stability of commonly used housekeeping genes in selected porcine tissues[J].BMC Research Notes,2011(4):441.
[11]Morgan S B,Frossard Jean-Pierre,Pallares F J,et al.Pathology and Virus Distribution in the Lung and Lymphoid Tissues of Pigs Experimentally Inoculated with Three Distinct Type 1 PRRS Virus Isolates of Varying Pathogenicity[J].Transboundary&Emerging Diseases,2014,63(3):285-295.
[12]任慧英,杨汉春,高云.猪繁殖与呼吸综合征病毒核酸在仔猪呼吸及繁殖系统动态分布的研究[J].中国兽医杂志,2002,38(2):3-4.
[13]荣福龙,刘永刚,孙广力,等.应用荧光定量RT—PCR方法检测PRRSV变异株在仔猪体内动态分布[J].中国预防兽医学报,2010,32(9):681-686.
[14]Tian K G,Yu X L,Zhao T Z,et al.Emergence of fatal PRRSV variants:unparalleled outbreaks of atypical PRRS in China and molecular dissection of the unique hallmark[J].PLo S One,2007,2(6):e526.
[15]刘永宏,韩春华,王凤龙,等.高致病性繁殖与呼吸综合征病毒在自然病例组织中的定位[J].中国兽医杂志,2009(6):24-25.