1.Assessing PCV2 antibodies in field pigs vaccinated with different porcine circovirus 2 vaccines using two commercial ELISA systems.
Min Kyoung SHIN ; Seung Hyun YOON ; Myung Hwui KIM ; Young Soo LYOO ; Seung Won SUH ; Han Sang YOO
Journal of Veterinary Science 2015;16(1):25-29
Porcine circovirus type 2 (PCV2) is the primary causative agent for post-weaning, multisystemic, wasting syndrome. Consequently, serologic detection of and vaccination against PCV2 are important for the swine industry. Among several serological tests, the enzyme-linked immunosorbent assay (ELISA) is commonly used to measure anti-PCV2 antibody levels. In the present study, we used two commercial ELISA systems to comparatively evaluate anti-PCV2 antibodies in field pigs treated with three different PCV2 vaccines. Among a total of 517 serum samples, the results of the two ELISAs were fully concordant for 365 positive and 42 negative samples, indicating 78.7% agreement. In addition, the Pearson coefficient (0.636) indicated a moderate correlation between data from the two ELISAs. Results from the farms with pigs vaccinated with the three different PCV2 vaccines demonstrated that most of the vaccinated animals underwent seroconversion. However, the increase and duration of antibody titers varied depending on the vaccine, the presence of maternal antibodies, and the vaccination program. PCV2 serologic status and anti-PCV2 antibody levels of herds from this study could be utilized to determine the best timing for vaccination and assessing vaccination compliance.
Aging
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Animals
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Antibodies, Viral/*blood
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Circovirus/*classification/immunology
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Enzyme-Linked Immunosorbent Assay/methods/*veterinary
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Female
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Porcine Postweaning Multisystemic Wasting Syndrome/blood/immunology/*prevention & control
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Republic of Korea/epidemiology
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Swine
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Swine Diseases/*prevention & control/virology
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Viral Vaccines/*immunology
2.Construction and identification of a recombinant PRRSV expressing ORF2 of porcine circovirus type 2.
Tingjie ZHANG ; Xing LIU ; Tao SUN ; Xuejiao ZHU ; Baochao FAN ; Juan BAI ; Ping JIANG
Chinese Journal of Virology 2015;31(1):65-73
Porcine reproductive and respiratory syndrome virus (PRRSV) and porcine circovirus type 2 (PCV2) are very two important pathogens that have coursed huge economic losses in swine production in worldwide. In this study,a vector pCMV-TJM containing the full-length cDNA clone of PRRSV attenuated strain TJM-F92 was firstly constructed by PCR method. Then a gene sequence containing Afl II/Mlu I e restriction enzyme sites and a transcription regulatory sequence for ORF6 (TRS6) was inserted be- tween ORF7 and 3'UTR, yielding a expression vector pCMV-TJM-TRS. Subsequently, a plasmid pCMV-TJM-Cap was constructed by cloning of PCV2 ORF2 gene into the unique sites Afl II /Mlu I of pCMV- TJM-TRS plasmid DNA. Then three recombinant PRRSV, rTJM, rTJM/TRS and rTJM/Cap, were rescued by transfection of pCMV-TJM, pCMV-TJM-TRS and pCMV-TJM-Cap into Marc-145 cells, respectively,and confirmed by the genome sequence, restriction enzyme digestion, Western Blot and IFA. They all had the molecular markers which was different from the parent virus. The growth characteristics of the rescued viruses were similar to that of parent virus. rTJM/Cap could also express efficiently PCV2 Cap protein in Marc-145 cells. At passage 8, it still had PCV2 ORF2 gene which examined by RT-PCR. It indicated that the full-length cDNA clone of PRRSV attenuated strain TJM-F92 and recombinant PRRSV rTJM/Cap expressing PCV2 Cap protein were successfully constructed. It made an important foundation for studying on the pathogenic mechanisms of PRRSV and PRRSV-PCV2 vaccine in the future.
Animals
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Capsid Proteins
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genetics
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immunology
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Cell Line
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Circoviridae Infections
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veterinary
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virology
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Circovirus
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classification
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genetics
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metabolism
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Gene Expression
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Open Reading Frames
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Porcine Reproductive and Respiratory Syndrome
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virology
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Porcine respiratory and reproductive syndrome virus
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genetics
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metabolism
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Recombination, Genetic
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Swine
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Swine Diseases
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virology
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Viral Vaccines
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genetics
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immunology