• Title, Summary, Keyword: porcine circovirus(PCV)

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Detection of viral pathogens and isolation of porcine circovirus 2 from postweaning multisystemic wasting syndrome-affected piglets (이유자돈 전신소모성증후군 이환 자돈에서의 바이러스성 원인체 검색 및 porcine circovirus 2 분리동정)

  • Park, Choi-Kyu;Kim, Hyun-Soo
    • Korean Journal of Veterinary Research
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    • v.44 no.4
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    • pp.561-569
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    • 2004
  • To detect viral agents and isolate porcine circovirus 2 (PCV2), 60 samples of lung and lymph node were collected from 5 to 12 week-old pigs that had showed clinical signs of postweaning multisystemic wasting syndrome (PMWS). Polymerase chain reactions (PCRs) were conducted to identify the viral pathogens including PCV1, PCV2, porcine parvovirus (PPV) and porcine reproductive and respiratory syndrome virus (PRRSV) that have been considered to be the causal agents of PMWS. Among 60 samples, PCV 2 was detected from 57 samples but no PCV 1 was detected. PRRSV and/or PPV were also detected from 27 (47.4%) samples and 1 (1.8%) sample of these 57 PCV 2-positive samples, respectively. Tissue homogenates were inoculated onto PCV-free PK-15 cell monolayers. Seven isolates were confirmed as PCV 2 by multiplex PCR, indirect immunofluorescence assay, and transmissible electron microscopy. These date suggest that PRRSV is a major cofactors causing PMWS in pigs that were infected with PCV2 in Korea.

Production and diagnostic applications of monoclonal antibodies against porcine circovirus (돼지 써코바이러스에 대한 단크론항체 생산 및 진단적 응용)

  • Kim, Kyung-Mi;Jeong, Ji-Hye;Min, Hong-Ki;Lee, Seung-Chul;Roh, In-Soon;Kang, Shien-Young
    • Korean Journal of Veterinary Research
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    • v.44 no.2
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    • pp.259-268
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    • 2004
  • Porcine circovirus type 2 (PCV-2) has been associated with various disease in pigs worldwide including postweaning multisystemic wasting syndrome (PMWS) and porcine dermatitis and nephropathy syndrome (PDNS). In this study, monoclonal antibodies (MAbs) against PCV were produced, characterized and applications of MAbs as diagnostic reagents were described. Spleen or lymph node cells from BALB/c mouse immunized respectively with PCV-1, PCV-2 or expressed PCV-2/ORF2 proteins in baculovirus were fused with SP2/0 myeloma cells using polyethylene glycol (PEG) and hybridoma cells producing PCV-1 or PCV-2-specific antibody were screened by an indirect immunofluorescence (IIF) test. A total of fifteen MAbs were produced against PCV. Six MAbs were PCV-1-specific and nine were PCV-2-specific. All PCV-1-specific MAbs reacted with only PCV-1 and all PCV-2-specific MAbs were reactive with only PCV-2 by IIF test. None of the MAbs was reactive with porcine reproductive and respiratory syndrome virus (PRRSV), porcine parvovirus (PPV), porcine rotavirus (PRV), and transmissible gastroenteritis virus (TGEV). Some PCV-2-specific MAbs recognized the PCV-2 infected porcine tissues by IIF or immunohistochemistry (IHC) assay. From this experiment, it was confirmed that MAbs produced in this study were PCV-specific and could be used as reliable diagnostic reagents for PCV-1/PCV-2 detection and differentiation.

Genetic variations in open reading frame 2 gene of porcine circovirus type 2 isolated in Korea during 2016-2017

  • Kim, Kiju;Choi, Jong-Young;Lyoo, Kwang-Soo;Hahn, Tae-Wook
    • Korean Journal of Veterinary Research
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    • v.58 no.3
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    • pp.143-146
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    • 2018
  • The capsid protein of porcine circovirus type 2 (PCV2) encoded by open reading frame 2 (ORF2) is important for neutralizing activity against PCV2 infection. This study investigated the heterogeneity of the ORF2 gene of PCV2 isolated in Korea during 2016-2017. The results revealed that PCV2d is currently the predominant genotype. Moreover, comparison of ORF2 from 17 PCV2 isolates revealed 88.3-100% homology at the nucleotide (deduced amino acid 86.3-100%) level. Interestingly, 61.5% (8/13) of the PCV2d isolates had glycine at position 210. These data provide a useful information for PCV2 epidemiology in Korea.

Porcine circovirus: detection of antibodies and virus antigen in Chungbdk area (Porcine circovirus에 대한 항체가 조사 및 바이러스 항원 확인)

  • 강신석;박재명;이종진;류재윤;최해연
    • Korean Journal of Veterinary Service
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    • v.24 no.2
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    • pp.127-132
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    • 2001
  • Porcine circoviruses(PCV) are the smallest nonenveloped DNA viruses containing a unique single-stranded circular genome. No recognized link was found between PCV infection of pig and disease. But the PCV consistently identified from postweaning multisystemic wasting syndrome(PMWS) and researches indicate that there are strong relationships between PCV and PMWS. Clinical signs were emaciation, dyspnea, high fever with normal appetite. Necropsy findings showed respiratory disease complex lesion and lymph node anomalities. An indirect-immunofluorescent antibody procedure was used to assay swine sera for the presence of PCV atibodies. Antibodies against PCV were found in an average of 20% of the samples tested. The PCV DNA was amplified from lymph nodes collected from pigs. PCV specific primers were successfully amplified PCV DNAs. Further studies are needed to determine the possible role this virus might have in disease.

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Genetic characterization of porcine circovirus 2 Korean isolates (Porcine circovirus 2 국내 분리주의 유전적 특성)

  • Park, Choi-Gyu;Lee, Kyoung-Ki;Kim, Hyun-Soo
    • Korean Journal of Veterinary Research
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    • v.44 no.4
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    • pp.571-579
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    • 2004
  • In order to obtain the genetic informations of the Korean isolates of porcine circovirus 2 (PCV2), nucleotide sequences of total genome of three isolates and open reading frame 2 (ORF2) of four isolates were determined and compared with those of other reference PCV2 isolates. Nucleotide sequences of 3 isolates showed over 99% homology with those of reference strain (GenBank accession no. AF027217). Point mutations were mainly determined on ORF2 regions but little on ORF1 regions. The patterns of pointmutated sites and nucleotide substitution on ORF2 regions were generally consistent between Korean isolates, and these mutated sites observed in Korean isolates were also relatively similar to those of foreign isolates. Phylogenetic analysis of nucleotide or amino acid sequences showed that there were minor branches consisting of three clusters; cluster of Korea, Canada and America, cluster of Spain and Taiwan, and the last cluster of French and China isolates. These results suggested that Korean PCV2s were probably originated from North America such as Canada or USA. The genetic informations obtained from this study could be useful for the research of diagnosis and pathogenecity of PCV2.

Application of Oral Fluid Sample to Monitor Porcine circovirus-2 Infection in Pig Farms (구강액을 이용한 양돈장의 Porcine circovirus-2 감염에 대한 모니터링)

  • Kim, Won-Il
    • Journal of Veterinary Clinics
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    • v.27 no.6
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    • pp.704-712
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    • 2010
  • Porcine circovirus-2 (PCV2) has been implicated in many clinical diseases/syndromes that are now referred to as PCV-associated diseases (PCVAD). Due to significant economic losses caused by PCVAD, many swine operations have launched extensive monitoring programs for PCV2. Traditional serum sampling is, however, rather expensive and laborious, hampering effective large scale pathogen surveillance. A field-based longitudinal study was conducted to assess the utility of pen-based oral fluid sample as an alternative to serum for herd PCV2 testing. Six pens (25 pigs/pen) at each of 3 different sites were used in the study. One oral fluid and 5 random serum samples per pen were collected at 3, 5, 8, 12, and 16 weeks of age, and the sera were pooled by pen for testing. All samples were tested for PCV2 by real-time PCR and for antibodies by indirect fluorescent antibody test (for both anti-PCV2 IgG and IgA) and 3 ELISA assays (blocking ELISA, indirect ELISA, and IgG/IgM sandwich ELISA). PCV2 DNA was detected in oral fluid samples sporadically until 8 weeks and in all pens at 16 weeks. PCV2-specific IgG was detected in oral fluid samples at 3 weeks and persisted until 5 to 8 weeks in all sites. Anti-PCV2 IgG and IgA were detectable in oral fluid samples collected at 16 weeks from all of the pens at 1 site. The detection of PCV2 and anti-PCV2 antibody in oral fluid samples correlated positively with results on pooled sera, suggesting that oral fluids can be a cost-effective alternative to serum for herd monitoring of PCV2 infection.

Application of PCR for diagnosis of porcine circovirus type 2 (돼지 써코바이러스 2형의 진단을 위한 PCR법 적용)

  • Park Hyo-Sun;Lee Hyo-Sang;Na Ki-Bok;Lee Kwan-Bok;Kang Su-Jung;Moon Sun-Hwa
    • Korean Journal of Veterinary Service
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    • v.29 no.1
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    • pp.1-8
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    • 2006
  • Porcine circovirus (PCV) is a small, nonenveloped virus that contains a single-stranded circular DNA genome of about 1.76 kb and belongs to the family circoviridae. The PCV-2 has been incriminated as the cause of post-weaning multisystemic wasting syndrome (PMWS) , an emerging disease in pigs. In the present study, a PCR assay was applied to detect PCV-2 in tissue samples. The presence of PCV-2 antigen in the porcine tissues was confirmed by indirect immunofluorescence (IIF) with PCV-2 specific monoclonal antibodies. And then DNA extracted from PCV-2 positive tissues was used as a template. One oligonucleotide primer suitable for PCR was selected from a published PCV-2 sequence (Genbank). Amplified PCR product was detected the same fragment lengths of 416 bp as a control. Based on these results, it was suggested that the PCR is a simple and sensitive method for support diagnostic purposes.

Rapid detection and quantification of porcine circovirus type 2 (PCV 2) DNA in Real-time PCR (Real-time PCR을 이용한 돼지써코바이러스 감염증 진단법 연구)

  • Kim, Eun-Gyeong;Hwang, Bo-Won;Lee, Jong-Min;Son, Byeong-Guk;Park, Ho-Jung;Kim, Tho-Kyoung
    • Korean Journal of Veterinary Service
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    • v.32 no.4
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    • pp.299-306
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    • 2009
  • Assay for the detection and quantification of porcine circovirus type 2 (PCV 2) with the real-time PCR were developed. TaqMan probe real-time using a set of primer/probe was developed for detection of PCV 2. In this study we applied real-time PCR assay to 320 samples, collected from pig farms. In 151 of 320 samples, PCV 2 DNA was detected by conventional PCR assay. All samples positive for PCV 2 DNA in conventional PCR assay were also positive in Real-time PCR assay, but 69 of 169 samples that tested negative for PCV 2 DNA in conventional assay were tested positive in TaqMan probe real-time PCR assay. The test of TaqMan probe real-time PCR resulted in detection and quantification limits of 101 copies per sample. TaqMan probe real-time PCR assay increased the number of samples in which PCV 2 was detected by 21%. TaqMan probe real-time PCR assay is very efficient method in contrast to the conventinal PCR, becoming increasingly important method for gene analysis.

Detection and genetic characterization of porcine circovirus 3 from aborted fetuses and pigs with respiratory disease in Korea

  • Kim, Seong-Hee;Park, Ji-Young;Jung, Ji-Youl;Kim, Ha-Young;Park, Yu-Ri;Lee, Kyoung-Ki;Lyoo, Young S.;Yeo, Sang-Geon;Park, Choi-Kyu
    • Journal of Veterinary Science
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    • v.19 no.5
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    • pp.721-724
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    • 2018
  • A novel porcine circovirus 3 (PCV3) was first detected in pigs showing porcine dermatitis and nephropathy syndrome, reproductive failure, and multisystemic inflammation in the USA. Herein, we report on PCV3 as a potential etiological agent of clinical signs, reproductive failure and respiratory distress on Korean pig farms, based on in situ hybridization, pathological, and molecular findings. Confirmation of the presence of PCV3 may increase co-infection with other causative agents of disease in Korean pig herds, indicating the need for further systemic investigation of pathogenicity and of multiple infections with PCV2 genotypes and bacteria, and the development of an effective PCV3 vaccine.

Development of serodiagnostic surface plasmon resonance imaging assay for the detection of antibodies to porcine circovirus type 2

  • Park, Chul;Kim, Bum-Seok;Kim, Yong-Hwan;Cho, Ho-Seong
    • Korean Journal of Veterinary Service
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    • v.34 no.1
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    • pp.1-4
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    • 2011
  • A surface plasmon resonance imaging (SPRI) assay was developed for measuring porcine circovirus type 2 (PCV2) antibody using a recombinant capsid protein as an antigen. The diagnostic potential of SPRI for detecting antibodies to the PCV2 capsid protein was compared with that of a conventional enzyme-linked immunosorbent assay (ELISA) using 70 pig serum samples taken from 6 pig farms. There was a strong positive correlation between the SPRI and ELISA (n = 70, r = 0.911, P<0.01). Therefore, this recombinant capsid protein can be used as an antigen for serological studies, and the SPRI, a label-free and high-throughput method, is expected to be a valuable tool in the serodiagnosis of PCV2 infection.