Detection of Procine Circovirus Type 2 by PCR Technique

Abstract

Abstract: 【Objective】A polymerase chain reaction for detecting PCV2 was established；【Mothed】Based on the gene sequence of PCV2, a pair of primers was designed.viral DNA from the sick tissues suspious of inclouding PCV2 extracted with phenol/chloroform as template, PCR technique was used; the special test of PCR detection was taken with the Hog choleravirus,Pseudorabies virus and Swine Parvovirus virus as control;the repetitive test of PCR detection was also taken.【Result】A size of 702 bp was amplified by PCR，and the PCR production was sequenced, sequence analysis showed that the sequence belonged to the genome of PCV2; the Hog choleravirus,Swine Pseudorabies virus and Swine Parvovirus virus were also detected by the PCR as control under the same condation with negative results; some sick tissues were detected for 5 times with high repetitive results.【Conclusion】The method of PCR detection was a sensitive, specific and reliable method for detecting PCV2, the sensitivity of PCR detection for PCV2 was up to 2×10-5ng／ml DNA of positive cell virus infected PCV2.

CLC Number:

S852.659

Wang Xianwen, Liu Xingyou, Liang Meilan, Zheng Yushu, Liu Liyan. Detection of Procine Circovirus Type 2 by PCR Technique[J]. Chinese Agricultural Science Bulletin, 2007, 23(9): 8-8.

References

[1] Tischer I,Rasch R,Tochtermann G. Characterization of papovavirus-and picornavirus-like particles in permanent pig kidney cell lines [J].Zbl Bakt Hyg,1974,226:153-167.
[2] Molnar T,Glavits R,Szeredi L,et al. Occurrence of porcine dermatitis and nephropathy syndrome in Hungary [J].Acta Vet Hung,2002,50(1):5-16.
[3] Elbers A R,Hunneman W A,Vos J H.et al. Increase in PDNS diagnoses in the Netherlands[J].Vet Rec,2000,147(11):311-315.
[4] Meehan B M, McNeilly F,McNair I, et al. Isolation and characterization of porcine circovirus 2 from cases of sow abortion and porcine dermatitis and nephropathy syndrome.[J].Arch Virol ,2001,146:835-842.
[5] Stevenson G. Distribution of porcine circovirus in neonatal pigs with congenital tremors.In:Proceedings of the CRWAD,1999,217-220.
[6] Drolet R,Larochelle R,Morin M, et al. Detection rates of porcine reproductive and respiratory syndrome virus, porcine circovirus type 2 and swine influenza virus in porcine proliferative and necrotizing pneumonia.[J]Vet Pathol, 2003.40:143-148.
[7] Larochelle R,Antaya M,Morin M, et al. Typing of porcine circovirus in clinical specimens by multiplex PCR. [J]. Virol Methods, 1999,80(1):69-75.
[8] Kim J, Chae C. Optimized protocols for the detection of porcine circovirus 2 DNA from formalin-fixed paraffin-embedded tissues using nested polymerase chain reaction and comparison of nested PCR with in situ hybridization. [J].J Virol Methods,2001,92:105-111.
[9] Kim J,Han DU,Choi, et al.Simultaneous detection and differentiation between porcine circovirus and porcine parvovirus in boar semen by multiplex seminested polymerase chain reaction[J].Vet Med Sci,2003,65(6):741-744.
[10] Fenaux M,Halbur PG,Gill M, et al.Genetic characterization of type 2 porcine circovirus(PCV-2) from pigs with postweaning muLtisystemic wasting syndrome in different geographic regions of North American and development of a differential PCR-restriction fragment length polymorphism assay to detect and differentiate between infections with PCV1 and PCV2[J].Clin Microbiol,2000,38(7):2494-2503.
[11] 郎洪武,张广川,吴发权,等.断奶猪多系统衰竭综合征血清抗体检测[J].中国兽医科技,2000,30(3):3-5.
[12] 踞春梅,陈焕春,刘正飞,等.应用在大肠杆菌中表达的猪2型圆环病毒ORF2蛋白建立一种ELISA诊断方法[J].畜牧兽医学报,2004,35(6),689-693.