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1.
猪圆环病毒3型(PCV3)作为一种新发现的圆环病毒,对养猪业的威胁持续升级。PCV3属于圆环病毒科、圆环病毒属,是一种无囊膜的环状DNA病毒。PCV3基因组由编码复制酶蛋白(Rep)和衣壳蛋白(Cap)的2个ORFs组成。其中,Cap蛋白在诱导宿主特异性免疫应答过程中发挥重要作用。2015年,美国学者Palinski等在患有猪皮炎肾病综合征和繁殖障碍母猪及其流产胎儿体内首次分离鉴定了PCV3。随后,中国、巴西、意大利、韩国、泰国、西班牙、丹麦、德国、瑞典和波兰等国家,陆续报道了PCV3感染病例。 相似文献
2.
《中国兽医学报》2016,(10):1658-1664
为对猪圆环病毒iDNA新型疫苗、基因组遗传变异以及基因组结构与功能等研究提供科学依据,运用相关生物信息学软件对猪圆环病毒2型(PCV2)贵州仁怀分离株(GZ-RH1)的基因组遗传特征进行了分析。结果表明,PCV2GZ-RH1株基因组结构与pmws(AF027217)株相比存在较大差异,其ORF2基因的第696位因碱基T的缺失而导致移码突变,致使其缺失了ORF8和ORF11,ORF5和ORF10终止密码子提前,其他ORF相对较为保守。PCV2GZRH1株各编码蛋白中除了ORF4和ORF9外,其余都属于亲水性蛋白,且ORF9含有1段信号肽序列。在ORF1和ORF2编码蛋白的磷酸化位点中,ORF1相对较为保守,而ORF2变异较大。ORF1~3所编码的蛋白除均具有各自的蛋白质家族域外,还预测到一些可能具有特殊功能的结构域,如ORF1编码蛋白的NACHT结构域和ORF2编码蛋白的精氨酸富集区。ORF2编码氨基酸位点熵值分析表明,ORF2蛋白氨基酸序列中包含2个高变区,即第53~91位和185~215位。通过构建进化树显示,GZ-RH1分离株系PCV2b亚型。对GZ-RH1株的密码子偏爱性分析表明,相对于大肠杆菌和酵母而言,昆虫细胞更适合于PCV2ORF2基因的体外表达。 相似文献
3.
为了解湖南猪群猪圆环病毒1型（PCV1）的遗传变异情况,以PCV1阳性DNA为模板,使用PCR方法分两段扩增PCV1全基因序列并进行拼接。结果共获得5个PCV1全基因组,其中3株全基因大小为1 759 bp,另外2株分别为1 758 bp和1 760 bp。5株全基因组同源性为98.4%~99.7%,与国内外的PCV1全基因组同源性为98.1%~99.7%;ORF2序列同源性为97.6%~99.6%,与国内外的PCV1 ORF2同源性为96.1%~99.9%。系统发育树显示,5株PCV1均属于同一分支,并显示出一定的地理差异,但差异较小。结果表明,湖南省流行的PCV1毒株基因较为稳定,分离株间差异较小。本研究为湖南省猪圆环病毒病防控及相关研究奠定了基础。 相似文献
4.
猪圆环病毒2型重组Cap蛋白在昆虫杆状病毒中的表达 总被引：3，自引：1，他引：3
猪圆环病毒2型(PCV2)基因组包含2个开放阅读框架(ORFs),其中ORF1编码病毒复制相关蛋白(Rep),ORF2编码病毒衣壳蛋白(Cap).为了在昆虫细胞表达Cap蛋白,本研究采用PCR扩增PCV2-ORF2编码基因,将PCR产物插入到昆虫杆状病毒转移载体上,经酶切反应及DNA序列分析得到验证.重组质粒与昆虫杆状病毒线性基因组混合,转染到昆虫细胞(Sf-21)进行基因重组,经3次病毒蚀斑克隆,获得高效表达Cap蛋白的重组杆状病毒,毒价可达1.28×108pfu/mL.采用SDS-PAGE凝胶电泳分析表明,重组Cap融合蛋白分子量为32.8 ku,占总蛋白含量的17.2%.免疫印迹试验分析表明,重组Cap蛋白与PCV2阳性血清产生特异性反应,证明该重组蛋白具有良好的免疫活性反应.本研究为进一步进行该病毒分子诊断、亚单位疫苗以及分子生物学等研究奠定了基础. 相似文献
5.
《中国预防兽医学报》2020,(5)
正猪圆环病毒2型(Porcine circovirus type 2,PCV2)属于圆环病毒科圆环病毒属的一员,是一类非常小的无囊膜的单股环状DNA病毒。PCV2病毒直径约为17 nm,基因组含有1 766 nt～1 768 nt核苷酸,主要包含4个开放阅读框架(Open reading frame, ORF), ORF1～ORF4,其中ORF2编码病毒的唯一结构蛋白,即衣壳蛋白(Capsid protein,Cap蛋白),与病毒感染和免疫息息相关。PCV2是一种 相似文献
6.
为原核表达猪圆环病毒2型(PCV2)的ORF3编码蛋白,本研究采用PCR方法以PCV2的CC株的基因组DNA为模板扩增ORF3基因,将其克隆到原核表达载体pET-32a(+)中,转化大肠杆菌BL21 (DE3),经IPTG诱导表达.SDS-PAGE结果显示,表达的重组蛋白约为31ku,并且以包涵体形式存在;western blot分析表明,ORF3重组蛋白能够与鼠抗His标签单克隆抗体发生特异性反应,表明原核表达的ORF3重组蛋白具有良好的反应原性. 相似文献
7.
从具有猪圆环病毒病临床症状的猪体内分离到1株PCV2华南分离株,并进行了全基因组序列测定。序列分析发现PCV2ORF2的变异原因主要是点突变,但也存在连续突变和缺失/插入突变,而PCV2 ORF1的变异均为点突变,变异程度很小。将此PCV2华南分离株的全基因组序列与GenBank上收录的18个PCV2毒株的全基因组序列做进化树分析,表明此PCV2华南分离株与国内PCV2分离株、欧洲株更接近,而与韩国、中国台湾、日本和美洲毒株则稍远,在基因进化树上,无法判定基因分支与地理位置是否有相关性。 相似文献
9.
猪圆环病毒Ⅱ型流行病学新特点及致病机理研究进展 总被引：1，自引：0，他引：1
猪圆环病毒是猪圆环病毒病的主要致病因素,该病给养猪业带来很大的经济损失。猪圆环病毒变异较快,是单链DNA病毒中最高的;易感猪主要通过消化道和呼吸道水平传播而感染,猪圆环病毒Ⅱ型(PCV2)人工或自然感染猪的排泄物、鼻腔、口腔和扁桃体拭子和患畜的尿液和粪便中均能检测到PCV2。断奶仔猪多系统衰竭综合征(PMWS)的发病主要与猪体内PCV2含量有关。PCV2特异抗体对猪群的PCV2感染具有保护作用。PCV2诱导猪继发性免疫缺陷,淋巴细胞缺失,影响PCV2感染结果的因素有:病毒、宿主、混合感染和免疫调节等。 相似文献
10.
生猪养殖过程中猪圆环病毒是危害生猪健康状况和生产性能的一种常见病毒,猪圆环病毒属于圆环病毒科圆环病毒属的DNA病毒,猪圆环病毒由于基因型的不同分为PCV1和PCV2两种,PCV1对猪只不具备致病性,而PCV2具有强致病性,通常会导致猪只出现混合性感染以及继发性感染的的情况,常见的混合性感染疾病包括猪瘟、猪蓝耳病、猪支原体肺炎以及猪细小病毒病、猪链球菌病等,给生猪的健康状况以及生产性能带来巨大的阻碍。下文将对生猪养殖过程中猪圆环病毒的危害以及净化措施进行介绍,旨在为生猪的健康养殖带来帮助。 相似文献
11.
Porcine circovirus type 2 (PCV2) is a disease caused by porcine circovirus, which can damage the host's immune system and exacerbate the clinical symptoms of many bacterial and viral infections.It has caused increasingly larger losses in the pig industry wordwide.The genomic DNA of PCV2 contains 11 open reading frames (ORFs) and at least seven potential ORFs-encoding proteins larger than 5 ku.However, only five virally encoded proteins, including Rep, Rep', Cap, ORF3 and ORF4, had been identified in PCV2 replication.In this review, we discussed the progress on structure and function of genomic DNA of PCV2 in order to provide insight into the scientific basis of the pathogenesis of PCV2 and prevent its infection. 相似文献
12.
猪圆环病毒1型和2型Rep蛋白在大肠杆菌中的表达与纯化 总被引：1，自引：0，他引：1
分别以猪圆环病毒1型基因组和重组质粒pCI-PCV2-ORF1为模板,利用PCR扩增了猪圆环病毒1型和2型的ORF1基因,随后克隆到pET-28a( )和pET-32a( )两种原核表达载体上。测序正确的重组质粒分别转化E.coli BL21(DE3),进行目的蛋白的诱导表达。SDS-PAGE和Western blot分析表明,猪圆环病毒1型和2型的ORF1均能在pET-28a和pET-32a中分别以重组蛋白His-Rep(40 Ku)和Trx-His-Rep(54 Ku)的形式表达。进一步纯化后测定重组蛋白的浓度,推算出猪圆环病毒1型和2型的His-Rep蛋白产量分别为34 mg/L菌液和14 mg/L菌液,Trx-His-Rep蛋白产量则为12 mg/L菌液和10 mg/L菌液。这些纯化的蛋白在Western blot中能够与猪抗猪圆环病毒2型阳性血清发生特异性反应,显示其具有良好的免疫学活性。本研究建立的猪圆环病毒重组Rep蛋白的原核表达系统及其纯化方法,为下一步开展Rep蛋白功能等相关研究奠定了基础。 相似文献
13.
Development of probes to differentiate porcine circovirus types 1 and 2 in vitro by in situ hybridization 总被引：8，自引：0，他引：8
Porcine circovirus type 1 (PCV1), a PK-15 cell line contaminant, and porcine circovirus type 2 (PCV2), associated with post-weaning multisystemic wasting syndrome (PMWS), are genetically and antigenically related. Several techniques have been developed to detect PCV, including in situ hybridization (ISH). Previously reported probes used for ISH may hybridize with both PCV1 and PCV2 nucleic acids. We attempted to produce probes for ISH that can detect and differentiate PCV2 from PCV1 in PCV-infected cells. Riboprobes were synthesized from the sense and antisense strands of both open reading frames 1 and 2 (ORF1 and ORF2) of PCV2. At 42 and 58 degrees C, the ORF1 antisense probe hybridized with nucleic acid from both PCV1- and PCV2-infected cells. At 58 degrees C, the ORF2 antisense probe hybridized with PCV2 nucleic acid but not with PCV1 nucleic acid. The ORF1 and ORF2 sense probes bound only with PCV2 nucleic acid. Both antisense strand probes produced stronger signals than the sense strand probes. The results showed that the PCV2 ORF1 antisense probe is the most likely probe to detect both PCV types while the ORF2 antisense probe is capable of discriminating between PCV1 and PCV2. 相似文献
14.
Seroprevalence of porcine circovirus type 2 in swine populations in Canada and Costa Rica 总被引：12，自引：1，他引：11 
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下载免费PDF全文 Qiang Liu Li Wang Philip Willson Brendan O'Connor Julia Keenliside Manuel Chirino-Trejo Ronald Melndez Lorne Babiuk 《Canadian journal of veterinary research》2002,66(4):225-231
Porcine circovirus (PCV) was recently divided into 2 antigenically distinct types that differ (65% amino acid identity) in the protein encoded by open reading frame 2 (ORF2). Porcine circovirus 1 is apparently non-pathogenic and, in contrast, PCV2 is associated with porcine multisystemic wasting syndrome (PMWS). Our objective was to determine the extent of exposure of normal pigs in Canada and Costa Rica to PCV2. Recombinant DNA techniques were used to produce an antigen from ORF2 of PCV2 that was suitable for the detection of antibody in swine sera. The presence of PCV2 nucleotide sequences was detected using polymerase chain reaction (PCR) techniques. Using these tests, specific antibody and nucleotide sequences were demonstrated in sera from a cohort of pigs during a PMWS outbreak. Antibody was detected in normal, healthy hogs slaughtered in Canada (82.4% of 386) and in Costa Rica (14.6% of 322). This is the first report indicating the presence of PCV2 in Latin America. More than 50% of these sera also contained PCV2 nucleotide sequence. Although these hogs were healthy when slaughtered, they were infected with PCV2 and may have previously been ill. The widespread occurrence of PCV2 in swine suggests that this virus is adapted to replication in porcine tissue. 相似文献
15.
Genotyping of porcine circovirus type 2 from a variety of clinical conditions in China 总被引：17，自引：0，他引：17
Genotypes of porcine circovirus type 2 (PCV2) in clinical tissue specimens collected from pigs of different region in China between 2001 and 2003 were analyzed by PCR and restriction fragment length polymorphism (RFLP) analysis of PCV2 genomic DNA encompassing the complete ORF2. The results showed that nine different genotypes (A-I) were identified and designated CHN-2A, CHN-2B, CHN-2C, CHN-2D, CHN-2E, CHN-2F, CHN-2G, CHN-2H and CHN-2I, respectively. Amongst the genotypes, 0.6% were CHN-2A (1/173), CHN-2B (1/173) and CHN-2C (1/173) RFLP profile; 2.3% were CHN-2F (4/173) and CHN-2G (4/173); 5.8% were CHN-2D (10/173); 8.6% were CHN-2E (15/173); 18.5% were CHN-2I (32/173) and 60.7% were CHN-2H (105/173). Therefore, our results suggest that CHN-2H is the dominant genotype of PCV2 prevailing in China. Sequence analysis revealed that ORF2 genes of different genotypic PCV2 exhibited the variation extent of 90.5-99.5% and 88-100% in nucleotide and amino acid respectively. The deduced amino acid sequences alignment of the capsid protein encoded by ORF2 of PCV2 presented that three major regions with greater heterogeneity existed at residues 57-90, 121-136 and 180-191 among nine genotypic PCV2. It was concluded that there exist variation in ORF2 genes of different genotypic PCV2 prevailing in China. 相似文献
16.
Non-radioactive digoxigenin (DIG)-labelled probes that can differentiate porcine circovirus (PCV) 1 from PCV2 in formalin-fixed, paraffin-wax-embedded tissues by in-situ hybridisation were developed. A 349 base pair (bp) DNA fragment from open reading frame (ORF) 1 of PCV1 and a 481 bp DNA fragment from ORF2 of PCV2 generated by polymerase chain reaction (PCR) were used as PCV1 and PCV2 probes, respectively. A specific DIG-labelled PCV1 DNA probe did not hybridise with PCV2-infected PK-15 cells and vice versa. From the 40 field cases with postweaning multisystemic wasting syndrome tested by in-situ hybridisation, 30 (75 per cent) cases were PCV2-positive only and 10 (25 per cent) cases were positive for both PCV1 and PCV2. PCV1 and PCV2 DNAS were detected mainly in the macrophages of lymph nodes and spleens. Positive cells typically exhibited a dark brown to black reaction product mainly in the cytoplasm but also occasionally in the nucleus. In-situ hybridisation together with the differential probes developed in the present study represent an additional tool capable of differentiating of both types of PCV in formalin-fixed, paraffin-wax-embedded tissues. 相似文献
17.
天津及周边地区猪圆环病毒2型流行情况调查及分离鉴定 总被引：1，自引：1，他引：0
为了调查天津及周边地区猪圆环病毒2型（porcine circovirus type 2,PCV2）的感染情况及其分子特征,本研究应用PCR方法对疑似PCV2感染病例开展调查,将阳性病料接种dulac细胞分离病毒,应用PCV2 ORF2基因特异性引物对分离的PCV2进行PCR扩增,克隆、测序后进行核苷酸序列同源性分析。结果显示,不同地区PCV2的阳性率为25.0%~38.1%,其中夏季的阳性率明显高于其他3个季节,达到44.0%;不同阶段猪群中,育肥猪群的PCV2阳性率最高,而哺乳母猪相对较低。经细胞分离得到24株PCV2,其ORF2序列与GenBank中PCV2序列的同源性为87.8%~99.2%,24株分离株之间的同源性为89.4%~100.0%;遗传进化分析显示,24个分离株归属于2个分支。对其中一株分离毒株（Y16155-1株）的体外增殖特性研究表明,第4代接种dulac细胞后6 h即可检出PCV2核酸,72 h病毒含量达到高峰,病毒滴度为10-4.3 TCID50/0.2 mL。本试验结果为进一步研究该地区PCV2分子流行病学及相关疾病的免疫防控奠定了基础。 相似文献
18.
Sinha A Lin K Hemann M Shen H Beach NM Ledesma C Meng XJ Wang C Halbur PG Opriessnig T 《Veterinary microbiology》2012,158(1-2):95-103
The objective of this study was to investigate cytokine expression and in vitro replication of porcine circovirus type 2 (PCV2) and porcine reproductive and respiratory syndrome virus (PRRSV) in pulmonary alveolar macrophages (PAMs) emphasizing PCV2 open-reading frame (ORF) origin (PCV2a or PCV2b) and PRRSV strain. Chimeric PCV2 viruses composed of different combinations of ORF1 and ORF2 of PCV2a or PCV2b (chimera PCV2a-2b and chimera PCV2b-2a) were constructed and five different PRRSV isolates were utilized: Type 1 (SD 01-08) or type 2 (NC16845b, VR-2332, MN-184, JA-142). PAMs were infected singularly or with combinations of PCV2b, PCV2a, chimera PCV2a-2b, and chimera PCV2b-2a, and one of the five PRRSV isolates. Real-time PCR was used to test PAMs (PCV2 mRNA) and supernatants (PRRSV RNA, PCV2 DNA, PCV2 mRNA) harvested at 24, 48, 72 and 96h post inoculation (hpi). Levels of IFN-γ, TNF-α and IL-10 were determined by quantitative ELISAs. PCV2 replication in PAMs was limited to groups inoculated with PCV2 strains containing ORF1 of PCV2a (PCV2a, chimera PCV2a-2b). Furthermore, in supernatants, PCV2 mRNA was only detected in groups coinfected with PRRSV regardless of strain at 48hpi supporting an enhancing effect of PRRSV on PCV2 infection. Changes in cytokine levels were minimal and associated with PRRSV strain for TNF-α. In summary, in vitro differences in PCV2 replication in PAMs inoculated with different PCV2-PRRSV combinations were independent of PCV2 ORF2 origin with minimal effects of concurrent PRRSV infection perhaps indicating that PCV2-specific changes in ORF1 may be more important than those in ORF2. 相似文献
19.
John Ellis Maria Spinato Choon Yong Keith West Francis McNeilly Brian Meehan Seamus Kennedy Edward Clark Steven Krakowka Gordon Allan 《Journal of veterinary diagnostic investigation》2003,15(4):364-368
Porcine circovirus 2 (PCV2) was first identified in high-health herds of domestic swine and was associated with a debilitating disease called postweaning multisystemic wasting syndrome (PMWS). Most subsequent studies have indicated that PCV2 infects only swine but there is little information on porcids other than improved breeds of domestic swine. Multisystemic disease was reported in a group of Eurasian wild boars raised under free-range conditions. Affected young pigs had pneumonia and enteritis and were cachectic. Porcine circovirus 2 was identified in affected tissue by immunohistochemistry and in situ hybridization, and a PCV2-like virus was isolated from pooled organs. The open reading frame (ORF2) of the isolated PCV2 had a 98.7% homology with the ORF2 of a reference PCV2 isolate. These diagnostic data indicate that PCV2 can infect and cause disease in Sus scrofa subspecies other than domestic swine. 相似文献