Author:
Zhou Shun,Han Si,Shi Jianli,Wu Jiaqiang,Yuan Xiaoyuan,Cong Xiaoyan,Xu Shaojian,Wu Xiaoyan,Li Jun,Wang Jinbao
Abstract
Abstract
Background
Porcine circovirus type 2 (PCV2) is the primary causative agent of the emerging swine disease known as postweaning multisystemic wasting syndrome (PMWS). Nowadays, polymerase chain reaction (PCR) is still the most widespread technique in pathogen detection. Loop-mediated isothermal amplification (LAMP), a novel nucleic acid amplification method developed in 2000, will possibly replace PCR in the field of detection. To establish a LAMP method for rapid detection of PCV2, two pairs of primers were designed specially from the open reading frame 2 (ORF2) sequences of PCV2. A LAMP method for rapid detection of PCV2 was established. To compare with PCR, sensitivity and specificity of LAMP were evaluated using the optimized reaction system. The LAMP products could be determined by agarose gel electrophoresis or adding SYBR Green I dye.
Results
The amplification of LAMP could be obtained at 63°C for 60 min. The detection limit was nearly 1 copy of DNA plasmid, more sensitive than PCR. There was no cross-reaction with porcine circovirus type 1 (PCV1), porcine pseudorabies virus (PRV) and porcine parvovirus (PPV) under the same conditions.
Conclusions
LAMP is an useful rapid detection method with high sensitivity and specificity for PCV2.
Publisher
Springer Science and Business Media LLC
Subject
Infectious Diseases,Virology
Reference19 articles.
1. Tischer I, Gelderblom H, Vettermann W, Koch M: A very small porcine virus with a circular single-stranded DNA. Nature. 1982, 295: 64-66. 10.1038/295064a0.
2. Allan GM, McNeilly F, Kennedy S, Daft B, Clarke EG, Ellis JA, Haines DM, Meehan BM, Adair BM: Isolation of porcine circovirus-like viruses from pigs with a wasting disease in the United States of America and Europe. J Vet Diagn Invest. 1998, 10: 3-10. 10.1177/104063879801000102.
3. Segalés J, Rosell C, Domingo M: Pathological findings associated with naturally acquired porcine circovirus type 2 associated disease. Vet Microbiol. 2004, 98: 137-149. 10.1016/j.vetmic.2003.10.006.
4. Darwich L, Segalés J, Domingo M, Mateu E: Changes in CD4+,CD8+,CD4+CD8+,and Immunoglobulin M-Positive Peripheral Blood Mononuclear Cells of Postweaning Multisystemic Wasting Syndrome-Affected Pigs and Age-Matched Uninfected Wasted and Healthy Pigs Correlate with Lesions and Porcine Circovirus Type 2 Load in Lymphoid Tissues. Clin and Diag Lab Immun. 2002, 9: 236-242.
5. Guo LJ, Lu YH, Wei YW, Huang LP, Liu CM: Porcine circovirus type 2 (PCV2): genetic variation and newly emerging genotypes in China. Virol J. 2010, 7: 273-10.1186/1743-422X-7-273.
Cited by
20 articles.
订阅此论文施引文献
订阅此论文施引文献,注册后可以免费订阅5篇论文的施引文献,订阅后可以查看论文全部施引文献