猪圆环病毒的研究进展

近年来断奶仔猪多系统衰竭综合征(PMWS)在全世界范围内呈不断流行和蔓延的趋势。猪圆环病毒Ⅱ型(PCV-2),目前已被世界各国学者确认为是引起PMWS的主要病原体,许多对PCV-2的研究也主要是针对PMWS的。本文就近年来对猪圆环病毒的病原及致病性、流行病学、诊断及防制等方面的研究进展作一综述。     

猪圆环病毒的研究进展

近年来断奶仔猪多系统衰竭综合征（PMWS）在全世界范围内呈不断流行和蔓延的趋势。猪圆环病毒Ⅱ型（PCV-2）,目前已被世界各国学者确认为是引起PMWS的主要病原体,许多对PCV-2的研究也主要是针对PMWS的。本文就近年来对猪圆环病毒的病原及致病性、流行病学、诊断及防制等方面的研究进展作一综述。     

猪圆环病毒PCR检测方法的建立

猪圆环病毒PCV属于圆环病毒科,根据国外已经报道的猪圆环病毒的序列,设计了下面的一对引物,上游引物和下游引物(upper:5'-TTTTTATCACTTCGTAATGGTTTTT-3'lower:5'-GGCCCCCTTCCTCCGTG-GATTGTTC-3'),提取DNA,对PCV-2进行PCR试验.然后对PCR产物进行琼脂糖凝胶电泳.结果显示,从美国进口的种毒中扩增出了长为1768bp的基因片段.本试验可以从病料或全血中检测出PCV,且该方法能区分PCV-1和PCV-2,快速、敏感、特异性强,适合于PCV的检测.     

猪圆环病毒研究进展

猪圆环病毒病是近些年来发现的一种新的动物病毒病,猪感染发病后,可引起断奶仔猪多系统衰竭综合症。本综述对该病毒的流行病学、生理学特性、致病机理、临床症状和诊断等方面进行阐述。     

猪圆环病毒感染与断奶仔猪多系统衰竭综合征

1974年 ,德国学者 Tischer等 [1 ] 从 PK- 15 (ATCCCCL31)细胞的污染病毒中检出一种形态学上与细小核糖核酸病毒相似的小球形病毒和乳多泡病毒样病毒粒子 ,并于1982年将其命名为猪圆环病毒 (porcine circovirus,PCV) [2 ] 。目前已知 PCV有 PCV- 1和 PCV- 2 2个血清型。PCV- 1来源于 PK- 15细胞 ,为非致病性 PCV(nonpathogenic PCV,np PCV ) [3 ] ;PCV- 2与断奶仔猪多系统衰竭综合征(postweaning m ultisystem ic wasting syndrom e,PMWS)密切相关 ,又称 PMWS相关 PCV[3 ] (PMWS- associated PCV,pmws- PCV)。 PM…     

猪圆环病毒感染

猪圆环病毒感染是近几年发现的猪的一种病毒性传染病，它可以引起仔猪先天性震颤和断奶猪多系统衰弱综合征。大量的研究证实，猪圆环病毒感染已成为对全球养猪业构成重大危害的传染病。     

猪圆环病毒的研究进展

猪圆环病毒(Porcine circovirus,PCV)是断奶仔猪多系统衰竭综合症(Postweaning pigs multi-systemic wasting syndrom,PMWS)的病原。PMWS是引起断奶仔猪死亡的一种重要传染病,严重威胁着我国的养猪业,由于迄今国内外尚无疫苗可进行免疫,也无理想的化学药物防治措施,PCV2感染的早期诊断和病猪的淘汰成为PMWS防治的重点。随着该病的不断出现,人们越来越注意到猪圆环病毒的危害性。为了更加全面、本质地认识该病毒,本文对其流行病学、理化特性、致病机理、诊断方法及分子生物学作了较为全面的综述。     

猪圆环病毒病的诊疗

<正>猪圆环病毒病又叫仔猪断奶多系统衰竭综合症,主要由2型圆环病毒及其他的病原体混合感染引起。主要症状是引起断奶仔猪皮肤发白,黄疸、剧烈下泻、呼吸困难、体重下降等。目前,该病是危害养猪业的头等大病,也是没有有效治疗方案和预防措施的一大传染病。笔者从事兽医工作二十余     

猪圆环病毒感染

猪圆环病毒(PorcineCircovirus,PCV)感染是近几年新发现的一种传染病,主要侵害哺乳仔猪和育肥猪.该病是由圆环病毒2型(PCV-2)所引起的疾病的总称,主要包括断奶仔猪多系统衰竭综合征(Postweaning Multisystemic Wasting Syndrome,PMWS)、猪繁殖障碍、猪呼吸道疾病、肠炎、增生性坏死性肺炎、猪皮炎与肾病综合症(Porcine Dermatitisand Nephropathy Syndrome,PDNS)、幼龄仔猪先天性震颤(Congenical Tremor,CT)等(杨汉春,2004;梁红虎等,2004).PCV还可以引起免疫抑制和免疫刺激,从而更容易与其他病毒、细菌继发和并发多种疾病.     

第七讲猪圆环病毒感染

猪圆环病毒可引起断奶仔猪多系统衰竭综合症（PMWS）猪间质性肺炎（IP），猪皮炎。肾病综合症（PDNS）以及圆环病毒引起的母猪繁殖障碍等，这些病总称为圆环病毒病（PCDV）。     

Detection and genotyping of porcine circovirus in naturally infected pigs by oligo-microarray

A rapid and reliable method for the identification of porcine circovirus (PCV) genotypes based on oligonucleotide microarray hybridization has been developed. The genotype-specific oligonucleotides (22-30 mer) immobilized on the surface of glass slides were selected to bind to the multiple target sites within the replication gene that are conserved among individual PCV genotypes. Cy5-labeled DNA targets were amplified in a PCR with primers common to both genotypes. The identification of PCV genotype was based on hybridization with several individual genotype-specific oligonucleotides. This approach combines the high sensitivity of PCR with the selectivity of DNA-DNA hybridization. The utility and feasibility of oligonucleotide microarray hybridization was evaluated by testing standard and 87 clinical isolates. Analysis of the specimens showed that this microarray-based method is capable of unambiguous identification of both genotypes and fivefold more sensitive than gel electrophoresis. Our results indicated that the oligonucleotide array is useful for the identification and discrimination of PCV from clinical isolates and specimens in a clinical laboratory.     

江西地方株猪圆环病毒感染的检测

对江西25个猪场有明显PMWS症状猪采集的病料,用PCR-RFLP(限制性片段长度多态性)技术检测猪圆环病毒1型、2型(PCV1、PCV2)。结果PCV2的阳性率为45％,PCV1的阳性率为40％,其中有3例仅为PCV2阳性,有1例仅为PCV1阳性,推测江西不仅存在PCV的感染,且感染率也较高。     

猪圆环病毒引发的断奶仔猪多系统衰竭综合征致病机理

断奶仔猪多系统衰竭综合征于1997年首次在加拿大西部发现,随后许多国家报道了本病的存在和发生,相继证实该病是由圆环病毒科(Circoviridae)猪圆环病毒2型(Porcinecircovirus 2,PCV-2)引起的。PMWS典型特征是全身性淋巴肿大。病毒感染后引发机体免疫抑制,可继发多种病原混合感染。经证实,PCV2感染对胸腺淋巴细胞的损伤在PMWS致病机理中发挥主导作用,巨噬细胞和树突状细胞浆内PCV-2检出率较高,但至今还不清楚哪些细胞亚群最先被PCV-2感染并提供了病毒复制的场所。PCV-2与细小病毒一样,需要依赖细胞生长S周期合成的聚合酶复制。许多研究者试图通过加强抗原刺激以提高进入S周期细胞数,却很难提高病毒在细胞培养中的增殖量。文章着重阐述PCV-2感染引发PMWS的致病机理以及未来需要开拓的研究方向。     

A review of porcine circovirus 2-associated syndromes and diseases

Clinical expression of porcine circovirus 2 (PCV2) infection in swine may result in several distinct syndromes and diseases including post-weaning multisystemic wasting syndrome (PMWS), porcine dermatitis and nephropathy syndrome (PDNS), reproductive failure, porcine respiratory disease complex, granulomatous enteritis, necrotizing lymphadenitis, and possibly exudative epidermitis. Association of PCV2 with congenital tremor in piglets is still controversial. The extent of the involvement of PCV2 in swine disease other than PMWS is currently poorly understood. This review concentrates on PCV-2-associated syndromes and diseases other than PMWS.     

Genetic diversity of porcine circovirus type 2 from pigs in Republic of Korea

Porcine circovirus type 2 (PCV2) is the causative agent of postweaning multisystemic wasting syndrome (PMWS). In this study, 38 PCV2 cases obtained from different pig farms with different health conditions in Republic of Korea were sequenced and analyzed. Phylogenetic analysis showed that our cases had a greater variation and the existence of two PCV2 groups with at least four subgroups (1A, 1C, 2D, and 2E). Most cases obtained from PMWS-affected herds were in group 1, whereas cases with no clinical signs compatible with PMWS (wasted non-PMWS) were included within group 2. Moreover, four cases from the wasted non-PMWS belonged to PCV2-group 1. Therefore, our results suggest that PCV2-group 1 is more related to PMWS than group 2.     

PCR detection of porcine circovirus type 2 (PCV2) DNA in blood, tonsillar and faecal swabs from experimentally infected pigs

PCV2 infection is now recognized as the major factor in the development of post-weaning multisystemic wasting syndrome (PMWS). In this study we evaluated the use of PCR to detect the presence of PCV2 DNA in blood, faecal and tonsillar swabs collected from 12 pigs experimentally infected with PCV2 and sampled at selected time points post-infection. The PCR results were evaluated together with the presence of PMWS typical histopathological lesions and the presence of PCV2 antigen. PCV2 DNA was present in the blood of all 12 infected pigs at the end of the experiment and faecal and tonsillar swabs of 11 of the 12 pigs. The rate of PCR-positive serum and plasma samples was significantly higher in four pigs that showed virological and pathological evidence of PMWS, than in infected pigs without evidence of disease. In conclusion this study confirms that PCR cannot substitute for the traditional methods used for diagnosis of PMWS, however, PCR amplification of PCV2 DNA from serum or plasma could be a useful tool to support an early diagnosis of PMWS in live animals.     

Genetic characterization of porcine circovirus type 2 in Republic of Korea

Porcine circovirus type 2 (PCV2) is a major causative agent of postweaning multisystemic wasting syndrome (PMWS). Sequence and phylogenetic analyses based on the ORF2 capsid protein gene fragment showed that field isolate in Republic of Korea (ROK), PCV2 YJK 0703, was closely related with the PCV2 Fh18 isolate. PCV2 YJK 0703 was genetically distinct and not related to previously reported ROK isolates. Therefore, genotypic variation exists among prevailing PCV2 in ROK. This result suggests that several PCV2 genotypes exist in Korean pig farms.     

Double in situ hybridization for simultaneous detection and differentiation of porcine circovirus 1 and 2 in pigs with postweaning multisystemic wasting syndrome

Double in situ hybridization using a digoxigenin-labelled porcine circovirus 1 (PCV1) and biotinylated PCV2 probe, was developed for the simultaneous detection and differentiation of PCV1 and PCV2 in formalin-fixed, paraffin-embedded tissues from pigs with postweaning multisystemic wasting syndrome. The combination of an alkaline phosphatase conjugated antidigoxigenin system with alkaline phosphatase conjugated streptavidin-biotin system allowed identification of PCV1 and/or PCV2. No evidence of cross-reaction was observed. Positive cells exhibited a red or dark brown reaction product for PCV1 and PCV2, respectively. Both PCV DNAs were observed mainly in the cytoplasm but occasionally in the nucleus. Co-localization of hybridization signal for both PCV1 and PCV2 was present in macrophages and multinucleated giant cells of the lymph node and spleen. This double-labelling technique for the differentiation between PCV1 and PCV2 is suitable for pathogenesis studies and diagnostic applications.     

Comparative analyses of humoral and cell-mediated immune responses upon vaccination with different commercially available single-dose porcine circovirus type 2 vaccines

The objective of this study was to compare the induction of humoral and cell-mediated immune responses by four commercially available single-dose porcine circovirus type 2 (PCV-2) vaccines. A total of 50 3-week-old piglets were assigned to five groups (10 pigs per group). Four commercial PCV-2 vaccines were administered according to the manufacturer's instructions and the piglets were observed for 154 days post vaccination (dpv). Inactivated chimeric PCV-1-2 vaccines induced higher levels of PCV-2-specific neutralizing antibodies (NA) and interferon-γ-secreting cells (IFN-γ-SC) in pigs than did the other three commercial PCV-2 vaccines. The proportions of CD4⁺ cells were significantly higher in animals vaccinated with inactivated chimeric PCV-1-2 and PCV-2 vaccines than in animals vaccinated with the two subunit vaccines. To our knowledge, this is the first comparison of humoral and cell-mediated immunity induced by four commercial single-dose PCV-2 vaccines under the same conditions. The results of this study demonstrated quantitative differences in the induction of humoral and cell-mediated immunity following vaccination.     

Efficacy of single dose of an inactivated porcine circovirus type 2 (PCV2) whole-virus vaccine with oil adjuvant in piglets

Background

Post-weaning multisystemic wasting syndrome (PMWS) associated with PCV2 is one of the most costly diseases currently faced by the swine industry. The development of effective vaccines against PCV2 infection has been accepted as an important strategy in the prophylaxis of PMWS.

Methods

In the present study, a PK-15 cell-adapted formalin-inactivated prototype vaccine candidate was prepared using a strain of PCV2 from China. Inactivation of the virus was accomplished using a standard formalin inactivation protocol. The protective properties of the inactivated PCV2 vaccine were evaluated in piglets. Ten 28-day-old pigs were randomly assigned to two groups, each with five. Group 1 was vaccinated intramuscularly with the inactivated virus preparation; Group 2 received sterile PBS as a placebo. By 28 days post-vaccination (DPV), Groups 1 and 2 were challenged intranasally and intramuscularly with 5 × 10⁷ TCID₅₀ of a virulent PCV2 isolate.

Results

The vaccinated pigs seroconverted to PCV2 and had high levels of serum antibodies to PCV2 at 28 days after vaccination, whereas the control pigs remained seronegative. No significant signs of clinical disease were recorded following the challenge with PCV2, but moderate amounts of PCV2 antigen were detected in most lymphoid organs of the control pigs. PCV2 was detected in two out of the five vaccinated pigs. Furthermore, pathological lesions and viremia were milder in the vaccinated group.

Conclusions

The obtained results indicate that the inactivated PCV2 virus vaccine with an oil adjuvant induce an immunological response in pigs that appears to provide protection from infection with PCV2. The vaccine, therefore, may have the potential to serve as a vaccine aimed to protect pigs from developing PMWS.