期刊论文详细信息
Virology Journal
A loop-mediated isothermal amplification assay for the visual detection of duck circovirus
Sisi Luo1  Qing Fan1  Zhiqin Xie1  Xianwen Deng1  Yaoshan Pang1  Jiabo Liu1  Guangyuan Zhao1  Zhixun Xie1  Liji Xie1 
[1] Department of Biotechnology, Guangxi Key Laboratory of Animal Vaccines and Diagnostics, Guangxi Veterinary Research Institute, 51 Youai North Road, 530001 Nanning, China
关键词: Visual detection;    Loop-mediated isothermal amplification;    Duck circovirus;   
Others  :  806020
DOI  :  10.1186/1743-422X-11-76
 received in 2014-01-15, accepted in 2014-04-25,  发布年份 2014
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【 摘 要 】

Background

Duck circovirus (DuCV) infection in farmed ducks is associated with growth problems or retardation syndromes. Rapid identification of DuCV infected ducks is essential to control DuCV effectively. Therefore, this study aims to develop of an assay for DuCV to be highly specific, sensitive, and simple without any specialized equipment.

Methods

A set of six specific primers was designed to target the sequences of the Rep gene of DuCV, and A loop-mediated isothermal amplification (LAMP) assay were developed and the reaction conditions were optimized for rapid detection of DuCV.

Results

The LAMP assay reaction was conducted in a 62°C water bath condition for 50 min. Then the amplification products were visualized directly for color changes. This LAMP assay is highly sensitive and able to detect twenty copies of DuCV DNA. The specificity of this LAMP assay was supported by no cross-reaction with other duck pathogens.

Conclusion

This LAMP method for DuCV is highly specific and sensitive and can be used as a rapid and direct diagnostic assay for testing clinical samples.

【 授权许可】

   
2014 Xie et al.; licensee BioMed Central Ltd.

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【 参考文献 】
  • [1]Hattermann K, Schmmitt C, Soike D, Mankertz A: Cloning and sequencing of duck circovirus (DuCV). Arch Virol 2003, 148:2471-2480.
  • [2]Fringuelli E, Scott AN, Beckett A, McKillen J, Smyth JA, Palya V, Glavits R, Ivanics E, Mankertz A, Franciosini MP, Todd D: Diagnosis of duck circovirus infections by conventional and real-time polymerase chain reaction tests. Avian Pathol 2005, 34:495-500.
  • [3]Chen C, Wang P, Lee M, Shien J, Shieh H, Ou S, Chen C, Chang P: Development of a polymerase chain reaction procedure for detection and differentiation of duck and goose circovirus. Avian Dis 2006, 50:92-95.
  • [4]Banda A, Galloway-Haskins RI, Sandhu TS, Schat KA: Genetic analysis of a duck circovirus detected in commercial Pekin ducks in New York. Avian Dis 2007, 51:90-95.
  • [5]Zhang X, Jiang S, Wu J, Zhao Q, Sun Y, Kong Y, Li X, Yao M, Chai T: An investigation of duck circovirus and co-infection in Cherry Valley ducks in Shandong Province, China. Vet Microbiol 2009, 133(3):252-256.
  • [6]Xie L, Xie Z, Zhao G, Liu J, Pang Y, Deng X, Xie Z, Fan Q: Complete Genome Sequence Analysis of a Duck Circovirus from Guangxi Pockmark Ducks. J Virol 2012, 86(23):13136-13136.
  • [7]Soike D, Albrecht K, Hattermann K, Schmitt C, Mankertz A: Novel circovirus in mulard ducks with developmental and feathering disorders. Vet Rec 2004, 154:792-793.
  • [8]Wan C, Fu G, Shi S, Cheng L, Chen H, Peng C, Lin S, Huang Y: Epidemiological investigation and genome analysis of duck circovirus in Southern China. Virol Sin 2011, 26(5):289-296.
  • [9]Liu SN, Zhang XX, Zou JF, Xie ZJ, Zhu YL, Zhao Q, Zhou EM, Jiang SJ: Development of an indirect ELISA for the detection of duck circovirus infection in duck flocks. Vet Microbiol 2010, 145(1):41-46.
  • [10]Wan C, Huang Y, Cheng L, Fu G, Shi SH, Chen H, Peng C, Lin F, Lin J: The development of a rapid SYBR Green I-based quantitative PCR for detection of Duck circovirus. Virol J 2011, 8:465. BioMed Central Full Text
  • [11]Halami MY, Nieper H, Müller H, Johne R: Detection of a novel circovirus in mute swans (Cygnus olor) by using nested broad-spectrum PCR. Virus Res 2008, 132:208-212.
  • [12]Notomi T, Okayama H, Masubuchi H, Yonekawa T, Watanabe K, Amino N, Hase T: Loopmediated isothermal amplification of DNA. Nucleic Acids Res 2000, 28:E63.
  • [13]Xie ZX, Tang Y, Fan Q, Liu JB, Pang YS, Deng XW, Xie ZQ, Peng Y, Xie LJ, Khan MI: Rapid detection of Group I avian adenoviruses by a Loop-mediated isothermal amplification. Avian Dis 2011, 55:575-579.
  • [14]Enomoto Y, Yoshikawa T, Ihira M, Akimoto S, Miyake F, Usui C, Suga S, Suzuki K, Kawana T, Nishiyama Y, Asano Y: Rapid diagnosis of herpes simplex virus infection by a loopmediated isothermal amplification method. J Clin Microbiol 2005, 43:951-955.
  • [15]Peng Y, Xie ZX, Lie JB, Peng YS, Deng XW, Xie ZQ, Xie LJ, Fan Q, Feng JX, Khan MI: Visual detection of H3 subtype avian influenza viruses by reverse transcription loop-mediated isothermal amplification assay. Virol Jour 2011, 8:1-10. BioMed Central Full Text
  • [16]Nemoto M, Imagawa H, Tsujimura K, Yamanaka T, Kondo T, Matsumura T: Detection of equine rotavirus by reverse transcription loop-mediated isothermal amplification (RT-LAMP). J Vet Med Sci 2010, 72:823-826.
  • [17]Rovira A, Abrahante J, Murtaugh M, Munoz-Zanzi C: Reverse transcription loop-mediated isothermal amplification for the detection of Porcine reproductive and respiratory syndrome virus. J Vet Diagn Invest 2009, 21:350-354.
  • [18]Xie Z, Fan Q, Liu J, Pang Y, Deng X, Xie Z, Xie L, Khan M: Reverse transcription loop-mediated isothermal amplification assay for rapid detection of Bovine Rotavirus. BMC vety res 2012, 8(1):133. BioMed Central Full Text
  • [19]Fan Q, Xie Z, Xie L, Liu J, Pang Y, Deng X, Xie Z, Peng Y, Wang X: A reverse transcription loop-mediated isothermal amplification method for rapid detection of bovine viral diarrhea virus. J Virol Methods 2012, 186(1–2):43-48.
  • [20]Xie Z, Xie L, Pang Y, Lu Z, Xie Z, Sun J, Deng X, Liu J, Tang X, Khan M: Development of a real-time multiplex PCR assay for detection of viral pathogens of penaeid shrimp. Arch Virol 2008, 153:2245-2251.
  • [21]Wang D, Xie X, Zhang D, Ma G, Wang X, Zhang D: Detection of duck circovirus in China: a proposal on genotype classification. Vet Microbiol 2011, 147(3):410-415.
  • [22]Tomita N, Mori Y, Kanda H, Notomi T: Loop-mediated isothermal amplification (LAMP) of gene sequences and simple visual detection of products. Nat Protoc 2008, 3:877-882.
  • [23]Woźniakowski G, Kozdruń W, Samorek-Salamonowicz E: Loop-mediated isothermal amplification for the detection of goose circovirus. Virol J 2012, 9(1):1-11. BioMed Central Full Text
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