中国动物传染病学报
中國動物傳染病學報
중국동물전염병학보
CHINESE JOURNAL OF VETERINARY PARASITOLOGY
2014年
2期
32-38
,共7页
程群%姜一峰%虞凌雪%王康%杨莘%李丽薇%高飞%于海%童武%童光志%周艳君
程群%薑一峰%虞凌雪%王康%楊莘%李麗薇%高飛%于海%童武%童光誌%週豔君
정군%강일봉%우릉설%왕강%양신%리려미%고비%우해%동무%동광지%주염군
高致病性猪繁殖与呼吸综合征病毒%PCR%分离%鉴定
高緻病性豬繁殖與呼吸綜閤徵病毒%PCR%分離%鑒定
고치병성저번식여호흡종합정병독%PCR%분리%감정
Highly pathogenic Porcine reproductive and respiratory syndrome virus (HP-PRRSV)%PCR%isolation%identification
2013年初上海某猪场保育猪群中出现高热、呼吸障碍等临床表现，导致大部分发病猪死亡，为了确定此次猪群中爆发疫病的病原，本研究从发病猪群中随机采集了15份血液样品，利用RT-PCR方法对本次疫病病原进行了检测。结果显示，有11份临床样品呈现猪繁殖与呼吸综合征病毒（Porcine reproductive and respiratory syndrome virus, PRRSV）强阳性，且与高致病性猪繁殖与呼吸综合征病毒（Highly pathogenic Porcine reproductive and respiratory syndrome virus, HP-PRRSV）对照大小相一致。随后从阳性样品中选取SHxx13/2013在Marc-145细胞中进行病毒的分离与鉴定，结果显示SHxx13/2013在细胞接种后第1代即出现明显的细胞效应（cytopathic effect, CPE），其病变特征与高致病性PRRSV强毒HuN4株一致。对SHxx13/2013第5代细胞分离毒进行RT-PCR和IFA等特异性鉴定，结果显示该毒株为PRRSV分离株，其蚀斑形态和生长特性与强毒HuN4株相似。分段克隆该毒株全长基因进行测序和序列分析，结果显示，新分离的流行毒SHxx13/2013株全长基因组为15319 bp，其Nsp2基因特征与HP-PRRSV一致，在第482位和第534~562位发生两处共30个氨基酸的不连续缺失，且该毒株与高致病性PRRSV代表毒株HuN4亲缘关系较近，全长基因的核苷酸同源性为99.6%。本研究结果表明新分离的SHxx13/2013株属于高致病性PRRSV分离株，据此我们推测今年年初上海某猪场爆发的疫情主要是由HP-PRRSV感染所致。
2013年初上海某豬場保育豬群中齣現高熱、呼吸障礙等臨床錶現，導緻大部分髮病豬死亡，為瞭確定此次豬群中爆髮疫病的病原，本研究從髮病豬群中隨機採集瞭15份血液樣品，利用RT-PCR方法對本次疫病病原進行瞭檢測。結果顯示，有11份臨床樣品呈現豬繁殖與呼吸綜閤徵病毒（Porcine reproductive and respiratory syndrome virus, PRRSV）彊暘性，且與高緻病性豬繁殖與呼吸綜閤徵病毒（Highly pathogenic Porcine reproductive and respiratory syndrome virus, HP-PRRSV）對照大小相一緻。隨後從暘性樣品中選取SHxx13/2013在Marc-145細胞中進行病毒的分離與鑒定，結果顯示SHxx13/2013在細胞接種後第1代即齣現明顯的細胞效應（cytopathic effect, CPE），其病變特徵與高緻病性PRRSV彊毒HuN4株一緻。對SHxx13/2013第5代細胞分離毒進行RT-PCR和IFA等特異性鑒定，結果顯示該毒株為PRRSV分離株，其蝕斑形態和生長特性與彊毒HuN4株相似。分段剋隆該毒株全長基因進行測序和序列分析，結果顯示，新分離的流行毒SHxx13/2013株全長基因組為15319 bp，其Nsp2基因特徵與HP-PRRSV一緻，在第482位和第534~562位髮生兩處共30箇氨基痠的不連續缺失，且該毒株與高緻病性PRRSV代錶毒株HuN4親緣關繫較近，全長基因的覈苷痠同源性為99.6%。本研究結果錶明新分離的SHxx13/2013株屬于高緻病性PRRSV分離株，據此我們推測今年年初上海某豬場爆髮的疫情主要是由HP-PRRSV感染所緻。
2013년초상해모저장보육저군중출현고열、호흡장애등림상표현，도치대부분발병저사망，위료학정차차저군중폭발역병적병원，본연구종발병저군중수궤채집료15빈혈액양품，이용RT-PCR방법대본차역병병원진행료검측。결과현시，유11빈림상양품정현저번식여호흡종합정병독（Porcine reproductive and respiratory syndrome virus, PRRSV）강양성，차여고치병성저번식여호흡종합정병독（Highly pathogenic Porcine reproductive and respiratory syndrome virus, HP-PRRSV）대조대소상일치。수후종양성양품중선취SHxx13/2013재Marc-145세포중진행병독적분리여감정，결과현시SHxx13/2013재세포접충후제1대즉출현명현적세포효응（cytopathic effect, CPE），기병변특정여고치병성PRRSV강독HuN4주일치。대SHxx13/2013제5대세포분리독진행RT-PCR화IFA등특이성감정，결과현시해독주위PRRSV분리주，기식반형태화생장특성여강독HuN4주상사。분단극륭해독주전장기인진행측서화서렬분석，결과현시，신분리적류행독SHxx13/2013주전장기인조위15319 bp，기Nsp2기인특정여HP-PRRSV일치，재제482위화제534~562위발생량처공30개안기산적불련속결실，차해독주여고치병성PRRSV대표독주HuN4친연관계교근，전장기인적핵감산동원성위99.6%。본연구결과표명신분리적SHxx13/2013주속우고치병성PRRSV분리주，거차아문추측금년년초상해모저장폭발적역정주요시유HP-PRRSV감염소치。
In early 2013, an acute disease characterized by high fever and respiratory problems in piglets outbroke on a pig farm in Shanghai and caused many deaths. To investigate the causative agent, a total of 15 serum samples were collected and viruses were examined in RT-PCR. Eleven out of of 15 clinical samples showed strong Porcine reproductire and respiratory syndrone virus (PRRSV) signal and the same PCR fragments as compared with highly pathogenic PRRSV (HP-PRRSV). One PRRSV strain was isolated from a PCR positive sample and designated as SHxx13/2013 strain. The SHxx13/2013 strain induced cytopathic effect (CPE) on MarC-145 cells at the first passage, which shared similar plaque morphology and growth kinetics with HuN4 strain. The complete genome of SHxx13/2013 strain was 15 319 bp. The deletion of total 30 amino acids occurred at positions 482 and 534-562 in Nsp2 gene, which was a common feature of HP-PRRSV. The complete genome of the SHxx13/2013 strain was 99.6%identity to that of HuN4 strain, indicating that it was close to HuN4. The results concluded that HP-PRRSV was the causative agent in pig population in Shanghai.
