A novel agent (hence termed as P2) was isolated from pig sera in China, which contained covalently bound circular genomic DNAs of 993 nucleotides. Sequence analyses indicated that the agent was closely related to the ...A novel agent (hence termed as P2) was isolated from pig sera in China, which contained covalently bound circular genomic DNAs of 993 nucleotides. Sequence analyses indicated that the agent was closely related to the porcine circovirus (PCV). The molecular clone of P2 was constructed subsequently and used for the following studies. Intracytoplasmic inclusions and intranuclear inclusions were only found in PK-15 cells transfected with the tandem dimer of P2 molecular DNA clone. Intracytoplasmic inclusions were round or irregular in shape and 0.1-0.4 μm in diameter, and intranuclear inclusions were electronically denser than intracytoplasmic inclusions and had two general shapes: round/small (0.1 μm in diameter) and hexagonal/large (0.5―1.4 μm in diameter). The inclusions were not membranously bound. The cells transfected with the tandem dimer of P2 molecular DNA clone were tested positive for P2 DNA at passages 5. The P2 antigen could be detected in both transfected and passaged PK-15 cells. This is the first report regarding the complete nucleotide sequence of a small DNA genome in a circovirus-like infectious agent in vitro.展开更多
Three pairs of primers P1/P2,P3/P4 and P5/P6 of porcine circovirus 2 were designed and synthesized to compare the sensitivity and specialization to amplify products of 404bp,647bp and 1773 bp.Optimum dilution of PK15 ...Three pairs of primers P1/P2,P3/P4 and P5/P6 of porcine circovirus 2 were designed and synthesized to compare the sensitivity and specialization to amplify products of 404bp,647bp and 1773 bp.Optimum dilution of PK15 cell genome infected with PCV 2 was achieved with 10-6ng,10-5ng to yield from the three pairs of primers.It was also confirmed that P1/P2 were 68.4% successful in detecting PCV2 in 57 swine samples from the province of Fujian,The other primers,P3/P4,P5/P6,were only 57.9% and 8.7% successful,respectively.We conclude that p1/p2 is the optimum primer pair for surveying incidents of PCV2 in pigs.展开更多
基金the State Key Basic Research Project (973 project) of China (Grant No. 2007CB116308)Planned Projects for Postdoctoral Research Funds of Jiangsu Province, China (Grant No. 5910602)Postdoctoral Funds of Jiangsu Academy of Agricultural Sciences, China (Grant No. 6510501)
文摘A novel agent (hence termed as P2) was isolated from pig sera in China, which contained covalently bound circular genomic DNAs of 993 nucleotides. Sequence analyses indicated that the agent was closely related to the porcine circovirus (PCV). The molecular clone of P2 was constructed subsequently and used for the following studies. Intracytoplasmic inclusions and intranuclear inclusions were only found in PK-15 cells transfected with the tandem dimer of P2 molecular DNA clone. Intracytoplasmic inclusions were round or irregular in shape and 0.1-0.4 μm in diameter, and intranuclear inclusions were electronically denser than intracytoplasmic inclusions and had two general shapes: round/small (0.1 μm in diameter) and hexagonal/large (0.5―1.4 μm in diameter). The inclusions were not membranously bound. The cells transfected with the tandem dimer of P2 molecular DNA clone were tested positive for P2 DNA at passages 5. The P2 antigen could be detected in both transfected and passaged PK-15 cells. This is the first report regarding the complete nucleotide sequence of a small DNA genome in a circovirus-like infectious agent in vitro.
文摘Three pairs of primers P1/P2,P3/P4 and P5/P6 of porcine circovirus 2 were designed and synthesized to compare the sensitivity and specialization to amplify products of 404bp,647bp and 1773 bp.Optimum dilution of PK15 cell genome infected with PCV 2 was achieved with 10-6ng,10-5ng to yield from the three pairs of primers.It was also confirmed that P1/P2 were 68.4% successful in detecting PCV2 in 57 swine samples from the province of Fujian,The other primers,P3/P4,P5/P6,were only 57.9% and 8.7% successful,respectively.We conclude that p1/p2 is the optimum primer pair for surveying incidents of PCV2 in pigs.
