Detection of PCV2 DNA by SYBR Green I-based quantitative PCR.

被引：48

作者：

Yang Z.Z. ^{[1
]}

Habib M. ^{[1
]}

Shuai J.B. ^{[1
]}

Fang W.H. ^{[1
]}

机构：

[1] Institute of Preventive Veterinary Medicine, Zhejiang University, Hangzhou

来源：

Journal of Zhejiang University SCIENCE B | 2007年 / 8卷 / 3期

关键词：

Porcine circovirus type 2 (PCV2); Real-time PCR; Sensitivity; S852.65;

D O I：

10.1631/jzus.2007.B0162

中图分类号：

学科分类号：

摘要：

We developed an assay for the detection and quantitation of porcine circovirus type 2 (PCV2) with the SYBR Green I-based real-time PCR. The real-time PCR provides a broad dynamic range, detecting from 10(3) to 10(11) copies of DNA per reaction. No cross-reactions were found in specimens containing PCV1. Because of the high sensitivity and specificity of the assay with a relatively rapid and simple procedure, real-time PCR can be used as a routine assay for the clinical diagnosis of PCV2 infection. In this study we applied real-time PCR assay to 80 clinical samples, collected from 40 pigs with postweaning multisystemic wasting syndrome (PMWS) and 40 healthy pigs in comparison with conventional PCR assay. In 56 of 80 samples, PCV2 DNA was detected by conventional PCR assay. All samples positive for PCV2 DNA in conventional PCR assay were also positive in real-time assay, and 12 of 24 samples that tested negative for PCV2 DNA in the conventional assay were tested positive in real-time PCR assay. Real-time PCR assay increased the number of samples in which PCV2 was detected by 15%. It is, therefore, considered to be a useful tool for the detection of PCV2.

引用

页码：162 / 169

页数：7

共 140 条

[1]

Allan G.M.(1995)Pathogenesis of porcine circovirus; experimental infections of colostrum deprived piglets and examination of pig foetal material Vet. Microbiol. 44 49-64

[2]

McNeilly F.(1998)Isolation of porcine circovirus-like viruses from pigs with a wasting disease in the USA and Europe J. Vet. Diagn. Invest. 10 3-10

[3]

Cassidy J.P.(1999)Isolation and characterisation of circoviruses from pigs with wasting syndromes in Spain, Denmark and Northern Ireland Vet. Microbiol. 66 115-123

[4]

Reilly G.A.(2000)Experimental infection of colostrum deprived piglets with porcine circovirus 2 (PCV2) and porcine reproductive and respiratory syndrome virus (PRRSV) potentiates PCV2 replication Arch. Virol. 145 2421-2429

[5]

Adair B.(2004)Quantitation of porcine circovirus type 2 isolated from serum/plasma and tissue samples of healthy pigs and pigs with postweaning multisystemic wasting syndrome using a TaqMan-based real-time PCR J. Virol. Methods 122 171-178

[6]

Ellis W.A.(2005)Real-time PCR for quantitation of porcine reproductive and respiratory syndrome virus and porcine circovirus type 2 in naturally-infected and challenged pigs J. Virol. Methods 124 11-19

[7]

McNulty M.S.(2004)Two amino acid mutations in the capsid protein of type 2 porcine circovirus (PCV2) enhanced PCV2 replication in vitro and attenuated the virus in vivo J. Virol. 78 13440-13446

[8]

Allan G.M.(2003)In vitro studies on the infection and replication of porcine circovirus type 2 in cells of the porcine immune system Vet. Immunol. Immunopathol. 94 149-161

[9]

McNeilly F.(2000)PCR detection and characterization of type-2 porcine circovirus Can. J. Vet. Res. 64 44-52

[10]

Kennedy S.(1997)Post-weaning multisystemic wasting syndrome (PMWS): preliminary epidemiology and clinical presentation Proc. Am. Assoc. Swine Pract. 28 503-214

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