Development of a colorimetric loop-mediated isothermal amplification assay for rapid and specific detection of Aves polyomavirus 1 from psittacine birds

• Published in: Journal of virological methods 2019-11, Vol.273, p.113687-113687, Article 113687
• Main Authors: Park, Min-Ji, Kim, Hye-Ryung, Chae, Ha-Gyeong, Lim, Da-Rae, Kwon, Oh-Deog, Cho, Kwang-Hyun, Park, Choi-Kyu
• Format: Article
• Language: English
• Subjects: Aves polyomavirus 1; Budgerigar fledgling disease; Loop-mediated isothermal amplification; Psittacine birds
• ISSN: 0166-0934; 1879-0984
• DOI: 10.1016/j.jviromet.2019.113687

•LAMP assay using HNB was developed for the rapid and visual detection of APyV.•The LAMP assay is simple, rapid, specific and sensitive.•The assay will be a valuable tool for the rapid diagnosis of APyV. A colorimetric loop-mediated isothermal amplification (LAMP) assay was developed for the rapid and specific detection of the T gene of Aves polyomavirus 1 (APyV), a causative agent of budgerigar fledgling disease (BFD) in psittacine birds. The amplification can be completed in 40 min at 60 °C, and the results can be visually detected by the naked eye using hydroxyl naphthol blue as a colorimetric indicator. The assay specifically amplified APyV DNA but not other viral and bacterial nucleic acids. The limit of detection of the assay was 5 × 102 DNA copies/reaction, which was comparable to those of previously reported conventional polymerase chain reaction assays. In the clinical evaluation, the LAMP results showed 100% concordance with those of the previously reported PCR assays with regard to specificity, sensitivity, and percentage of overall agreement, with a kappa value of 1.0. These results indicate that the developed LAMP assay will be a valuable tool for the rapid, sensitive and specific detection of APyV from BFD-suspected psittacine bird samples even in resource-limited laboratories.