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Open Access Highly Accessed Methodology

A loop-mediated isothermal amplification assay for the visual detection of duck circovirus

Liji Xie, Zhixun Xie*, Guangyuan Zhao, Jiabo Liu, Yaoshan Pang, Xianwen Deng, Zhiqin Xie, Qing Fan and Sisi Luo

Author Affiliations

Department of Biotechnology, Guangxi Key Laboratory of Animal Vaccines and Diagnostics, Guangxi Veterinary Research Institute, 51 Youai North Road, 530001 Nanning, China

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Virology Journal 2014, 11:76  doi:10.1186/1743-422X-11-76

Published: 29 April 2014

Abstract

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.

Keywords:
Duck circovirus; Loop-mediated isothermal amplification; Visual detection