Development of a Loop-mediated Isothermal Amplification Method for Rapid Detection of Beak and Feather Disease Virus in Parrots

Background: There are no effective antiviral treatments for Beak and Feather Disease virus (BFDV); thus, rapid diagnosis is critical for effective control of the disease. Recent development of a novel Loop-Mediated Isothermal Amplification (LAMP) technique that amplifies nucleic acids rapidly with high specificity and sensitivity under isothermal conditions has overcome some of the deficiencies of nucleic acid-based diagnostic tests and has made on-site diagnosis possible.

Methods and finding: We established a LAMP method for rapid detection of BFDV using 2 pairs of primers that were designed from BFDV and compared its sensitivity and specificity with PCR. Amplification by LAMP was optimal at 63°C for 60 min. The detection limit was nearly 3.5 fg of BFDV DNA— as sensitive as PCR. There was no cross-reaction with porcine circovirus type 2 (PCV2), pigeon circovirus (PiCV) or avian polyomavirus under the same conditions. The assay also successfully detected BFDV DNA in the tissues of infected parrots.

Conclusion: This is the first report indicating that LAMP is a valuable, rapid method of detecting BFDV with high sensitivity and specificity.

Author(s):

Yi Hsuan Kuo, Shinn Shyong Tsai, Hung Jen Liu and Kuo Pin Chuang

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