• Title/Summary/Keyword: Porcine circovirus-2

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Detection of porcine circovirus 2, porcine reproductive and respiratory syndrome virus and Mycoplasma hyopneumoniae from swine lungs with lesions by PCR (돼지 폐렴병변에서 PCR을 이용한 써코바이러스 2, 돼지생식기호흡기증후군, 마이코플라즈마 폐렴 감염실태 조사)

  • Chu, Keum-Suk;Kang, Mi-Seon;Jo, Young-Suk;Lee, Jeong-Won
    • Korean Journal of Veterinary Service
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    • v.31 no.1
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    • pp.71-77
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    • 2008
  • Today swine respiratory disease is one of the most important diseases because of its economic losses and severe infection nationwide, and swine society as well as veterinary service are trying to prevent the diseases in Korea. This study would like to obtain some information useful for the control of the diseases. A total of 174 lung specimens with lesion consisted of 3 sorts; 60 were collected from nursey pigs requested for diagnostic service from March of 2006 to October of 2007, 58 finishing pigs and 56 sows were selected from slaughterhouse from September to November 2007. In the detection test of pathogens by PCR, porcine circovirus 2 (PCV2), porcine reproductive and respiratory syndrome virus (PRRSV) and Mycoplasma hyopneumoniae were positive in 95.4%, 31.6%, and 20.1%, respectively. Double infection rate with PCV2 and PRRS was 30.4%, PCV2 and M hyopneumoniae was 19.5%, triple infection with PCV2, PRRS and M hyopneumoniae was 5.7%, respectively.

Prevalence and Detection of Porcine Circovirus 2 in Aborted Fetuses and Stillborn Piglets

  • H, Yoon-chul;Chae, Chan-hee
    • Proceedings of the Korean Society of Veterinary Pathology Conference
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    • 2003.10a
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    • pp.31-31
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    • 2003
  • Porcine circovirus (PCV) is the smallest virus that replicates autonomously in mammalian cells. PCV2 is recongized as the casual agent of postweaning multisystemic wasting syndrome (PMWS).[1] Although the role of PCV2 in reproductive failure requires further clarification, there have been several reports of PCV2-associated reproductive failure. Consistent clinical signs of affected farms include elevated abortion, stillbirths and fetal mummification. The objective of this study was to determine the prevalence of PCV2 in aborted fetuses and stillborn piglets. Second objective was to determine the distribution of PCV2 DNA and antigen in aborted fetuses and stillborn piglets by in situ hybridisation and immunohistochemistry. (omitted)

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Interaction between host cell proteins and open reading frames of porcine circovirus type 2

  • Si-Won Park;In-Byung Park;Seok-Jin Kang;Joonbeom Bae;Taehoon Chun
    • Journal of Animal Science and Technology
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    • v.65 no.4
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    • pp.698-719
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    • 2023
  • Postweaning multisystemic wasting syndrome (PMWS) is caused by a systemic inflammation after porcine circovirus type 2 (PCV2) infection. It was one of the most economically important pathogens affecting pig production worldwide before PCV2 vaccine was first introduced in 2006. After the development of a vaccine against PCV2a type, pig farms gradually restored enormous economic losses from PMWS. However, vaccine against PCV2a type could not be fully effective against several different PCV2 genotypes (PCV2b - PCV2h). In addition, PCV2a vaccine itself could generate antigenic drift of PCV2 capsid. Therefore, PCV2 infection still threats pig industry worldwide. PCV2 infection was initially found in local tissues including reproductive, respiratory, and digestive tracks. However, PCV2 infection often leads to a systemic inflammation which can cause severe immunosuppression by depleting peripheral lymphocytes in secondary lymphoid tissues. Subsequently, a secondary infection with other microorganisms can cause PMWS. Eleven putative open reading frames (ORFs) have been predicted to encode PCV2 genome. Among them, gene products of six ORFs from ORF1 to ORF6 have been identified and characterized to estimate its functional role during PCV2 infection. Acquiring knowledge about the specific interaction between each PCV2 ORF protein and host protein might be a key to develop preventive or therapeutic tools to control PCV2 infection. In this article, we reviewed current understanding of how each ORF of PCV2 manipulates host cell signaling related to immune suppression caused by PCV2.

UNG-based direct polymerase chain reaction (udPCR) for the detection of porcine circovirus 2 (PCV2) (UNG 기반 direct polymerase chain reaction (udPCR)을 이용한 돼지 써코바이러스 2형 진단법)

  • Kim, Eun-Mi;Park, Choi-Kyu
    • Korean Journal of Veterinary Service
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    • v.37 no.4
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    • pp.253-261
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    • 2014
  • Porcine circovirus disease (PCVD) is a major problem of swine industry worldwide, and diagnosis of PCV2, causal agent of PCVD, has been doing in clinical laboratories of pig disease by polymerase chain reaction (PCR) methods. But the PCR analyses have a serious problem of misdiagnosis by contamination of DNA, in particular, from carryover contamination with previously amplified DNA or extracted DNA from field samples. In this study, an uracil DNA glycosylase (UNG)-based direct PCR (udPCR) without DNA extraction process and DNA carryover contamination was developed and evaluated on PCV2 culture and field pig samples. The sensitivity of the udPCR combined with dPCR and uPCR was same or better than that of the commercial PCR (cPCR) kit (Median diagnostics, Korea) on PCV2-positive serum, lymph node and lung samples of the pigs. In addition, the udPCR method confirmed to have a preventing ability of mis-amplification by contamination of pre-amplified PCV2 DNA from previous udPCR. In clinical application, 170 pig samples (86 tissues and 84 serum) were analysed by cPCR kit and resulted in 37% (63/170) of positive reaction, while the udPCR was able to detect the PCV2 DNA in 45.3% (77/170) with higher sensitivity than cPCR. In conclusion, the udPCR developed in the study is a time, labor and cost saving method for the detection of PCV2 and providing a preventing effect for DNA carryover contamination that can occurred in PCR process. Therefore, the udPCR assay could be an useful alternative method for the diagnosis of PCV2 in the swine disease diagnostic laboratories.

Efficient Production of Porcine Circovirus Type 2 Capsid Protein using Baculovirus

  • Lee, Jun-Beom;Bae, Sung-Min;Kim, Hee-Jung;Lee, Won-Woo;Heo, Won-Il;Shin, Tae-Young;Choi, Jae-Bang;Woo, Soo-Dong
    • International Journal of Industrial Entomology and Biomaterials
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    • v.24 no.1
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    • pp.23-27
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    • 2012
  • Porcine circovirus type 2 (PCV2) is a single-stranded circular DNA virus associated with Postweaning multisystemic wasting syndrome (PMWS), which is considered to be an important infectious swine viral disease. PCV2 capsid protein encoded by ORF2 is a structural protein and expected as the high immunogenicity protein. In this study, we generated recombinant baculovirus containing ORF2 of PCV2 and analyzed the optimal conditions for the production of capsid protein in insect cell. Production and status of recombinant capsid protein in insect cell were confirmed by SDS-PAGE and Western blot analysis using His tag antibody and anti-PCV2 serum. The yield of recombinant capsid protein was high like as shown visible on SDS-PAGE. Optimal multiplicity of infection (MOI) and infection time of recombinant virus were determined as 5 MOI and 4 days, respectively. ORF2 is known to have N-linked glycosylation site, but we couldn't detect the glycosylation of recombinant protein in insect cells.

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.

Correlation between Disease Prevalence and Production Performance in Korean Swine Farms (양돈 생산성에 따른 주요 질병 분포 조사)

  • Jung, Ho-Kyoung;SunWoo, Sun-Young;Lyoo, Young-S.
    • Journal of Veterinary Clinics
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    • v.28 no.4
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    • pp.415-421
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    • 2011
  • Currently, various diseases reside in Korean swine farms and affect production performance of the farms greatly. These damages from disease are further aggravated by the concurrent infection of other disease. In this study, y investigating the distribution of porcine reproductive and respiratory syndrome virus (PRRSV), porcine circovirus type 2 (PCV2), Salmonella spp. in farms, correlation between the damage and the prevalence of disease was analyzed. Ten selected Korean swine farms that uses PCV2 vaccine were tested for presence of antibody and antigen of PRRSV, PCV2, Salmonella spp. per ages of pigs, 4weeks, 7weeks, 11weeks and 17weeks, respectively. The results were analyzed by dividing the farms in to groups with MSY above 19, and that with MSY below 19. Then calculating the distribution of disease each ages of pigs. Farms with MSY below 19 showed high prevalence of disease by PRRSV, PCV2 and Salmonella spp.. In this group, the detection rate of PCV2 and Salmonella spp. was increased by the activation/viremia of PRRSV in the young ages of pigs. The results are proved that the correlation between disease prevalence and production performance in Korean swine farms were very significant. The prevalence of PRRSV is more important index which influence to the productivity in current prevalence of diseases.

Pathogenesis and pathogenicity for the porcine circovirus 2 Korean isolates in weaned pigs (국내 분리 porcine circovirus 2의 이유자돈에 대한 병원성 시험연구)

  • Roh, In-Soon;Lee, Kyung-Woo;Kim, Jae-Hoon;Han, Jeong-Hee;Jean, Young-Hwa
    • Korean Journal of Veterinary Research
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    • v.47 no.2
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    • pp.175-185
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    • 2007
  • This study was carried out to investigate the pathogenesis and pathogenicity of the porcine circovirus type 2 (PCV2) Korean isolate from weaned pigs. Twenty four weaned pigs, PCV2, porcine reproductive and respiratory syndrome virus (PRRSV) and porcine parvovirus (PPV) antibodies free, were allocated to 4 groups (n = 6). Six pigs were inoculated intranasally with PCV2 alone, 6 with PCV2 and PRRSV, 6 with the combined PCV2/PRRSV/PPV inoculum, and 6 were remained as a uninoculated negative control. Pigs were killed 3 and 6 weeks after inoculation and tissue samples examined for gross and microscopic lesions and for the presence of PCV2 antigens and nucleic acids. Experimentally inoculated pigs were evaluated for 3 considerations: 1. development of postweaning multisystemic wasting syndrome (PMWS), 2. distribution of viral antigens by immunohistochemistry and polymerase chain reaction (PCR), and 3. cytokine mRNA levels in lymph nodes. Pigs inoculated with PCV2/PRRSV/PPV showed typical clinical signs, gross findings, and histopathologic characteristics of PMWS. In the PCV2/PRRSV/PPV inoculated group, the PCV2 antigen was widely distributed in various parenchymal organs such as brain, spinal cord, tonsil, lymph nodes, lung, heart, liver, kidney, spleen, and peyer's patch. Lymph node mRNA expression of IL-$1{\alpha}$, IL-2R and IL-8 was determined by real-time PCR. The pigs of PCV2/PRRSV and PCV2/PRRSV/PPV inoculation group, the mRNA expression was characterized by a decrease of IL-$1{\alpha}$, IL-2R and IL-8. The decrease of cytokine mRNA represent the state of T cell immuno-suppression in pig, and nicely support the evidence for the impairment of immune system in pigs with PMWS. In conclusion, PCV2 infection and some additional infectious causes such as PRRSV and/or PPV are warranted for the presence of PMWS in weaned pigs in Korea.

Visual detection of porcine circovirus 2 by loop-mediated isothermal amplification (LAMP) with hydroxynaphthol blue dye (육안 판독 등온증폭법을 이용한 돼지 써코바이러스 2형 신속 진단법)

  • Kong, Ho-Chul;Kim, Eun-Mi;Jeon, Hyo-Sung;Kim, Ji-Jung;Kim, Hee-Jung;Park, Yu-Ri;Kang, Dae-Young;Kim, Young-Hwa;Park, Jun-Cheol;Lee, Chang-hee;Yeo, Sang-Geon;Park, Choi-Kyu
    • Korean Journal of Veterinary Service
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    • v.38 no.3
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    • pp.145-153
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    • 2015
  • In this study, we developed a loop-mediated isothermal amplification (LAMP) with hydroxynaphtol blue dye (HNB) for rapid and direct visual detection of porcine circovirus 2 DNA with high sensitivity and specificity. The LAMP was completed in 40 min at $63^{\circ}C$, and the results of the LAMP can be confirmed by naked eye without any detection process. The sensitivity of the LAMP was 10-fold higher than that of the commercial PCR (cPCR) and real time PCR (rPCR) previously reported. In clinical application, the PCV2 detection rate of the LAMP was the same on porcine tissue samples (75.0%, 36/48) between porcine blood samples (75.0%, 39/52). The PCV2 detection rate (75.0%) of LAMP was higher than those of the cPCR and rPCR (67.3%, 35/52) in blood samples. In conclusion, the LAMP developed in the study could be an useful alternative method for the detection of PCV2 in the swine disease diagnostic laboratories.

Detection of etiological agents of proliferative and necrotizing pneumonia in pigs in Jeju (제주지역 돼지에서 증식성 괴사성 폐렴의 원인체 검출)

  • Kim, Jae-Hoon;Jung, Ji-Youl;Yang, Hyoung-Seok;Kim, Jae-Hoon
    • Korean Journal of Veterinary Service
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    • v.45 no.1
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    • pp.63-69
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    • 2022
  • Proliferative and necrotizing pneumonia (PNP) is a form of interstitial pneumonia that occurs in post-weaning pigs. In this study, we investigated the presence of swine influenza virus (SIV), porcine reproductive and respiratory syndrome virus (PRRSV), porcine circovirus type 2 (PCV2) and Aujeszky's disease virus (ADV) in PNP lesions in Jeju pigs. Based on the histopathologic criteria for PNP, a total of 50 cases were selected in Jeju pigs between 2008 and 2010. Coupled with histopathological examinations, the presence of ADV and SIV by polymerase chain reaction (PCR) and reverse transcription PCR (RT-PCR) and PRRSV and PCV2 by immunohistochemical (IHC) methods were investigated. Based on the PCR and RT-PCR methods, ADV and SIV nucleic acids were not detected in all cases. According to IHC, PRRSV was detected in 38 of the 50 cases examined (76%) and PCV2 in 25 cases (50%). PRRSV or PCV2 were detected in 19 (38%) or 6 (12%) cases, respectively. Both PRRSV and PCV2 were identified in other 19 cases (38%). Antigens of PRRSV and PCV2 were commonly observed in the cytoplasm of macrophages and clusters of necrotic cells in alveolar cavities. The results of the present study demonstrate that PRRSV is predominantly associated with PNP in Jeju pigs. Co-infection with PRRSV and PCV2 may enhance the severity of PNP lesions in affected pigs.