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Updated: Jul 2, 2026
Rapid Diagnosis of Avian Influenza Virus in Wild Birds: Use of a Portable rRT-PCR and Freeze-dried Reagents in the Field
Published on: August 2, 2011
Development of a rapid recombinase polymerase amplification assay for avian polyomaviruses
Jiaqian Rao1, Qian Wang1, Mingji Li1
1State Key Laboratory of Swine and Poultry Breeding Industry & Heyuan Branch, Guangdong Provincial Laboratory of Lingnan Modern Agricultural Science and Technology, College of Animal Science, South China Agricultural University, Guangzhou 510642 PR China; Key Laboratory of Animal Health Aquaculture and Environmental Control, Guangdong, Guangzhou 510642 China; South China Collaborative Innovation Center for Poultry Disease Control and Product Safety, Guangzhou 510642 China; Guangdong Provincial Key Lab of AgroAnimal Genomics and Molecular Breeding, College of Animal Science, South China Agricultural University, Guangzhou 510642 China; Guangdong Engineering Research Center for Vector Vaccine of Animal Virus, Guangzhou 510642 China.
None:
Budgerigar fledgling disease, caused by avian polyomavirus (APV), is an immunosuppressive condition in budgerigars that typically manifests as abdominal swelling, absence of down feathers on the dorsal and ventral regions, subdermal bleeding in nestlings, and high mortality. A new APV detection relied on real-time quantitative recombinase polymerase amplification (real-time RPA) targeting the large T antigen gene using specific probe and primers. Detection can be accomplished within 25 minutes at 41°C, with this detection threshold of 2.56 × 102 copies/μL. The real-time RPA assay specific detected APV and did not show cross-reactivity with avian infectious bronchitis virus (IBV), psittacine beak and feather disease virus (PBFDV), newcastle disease virus (NDV) and H9N2 avian influenza virus(H9N2). Intra-assay repeatability testing showed that the coefficients of variations (CVs) were all below 16%, while inter-assay repeatability testing showed that the CVs were all below 12%. In 63 parrot cases suspected of APV-associated mortality, tissue specimens were examined using real-time RPA alongside qPCR assays. Both detection methods showed identical positive rates, indicating complete agreement in positive/negative classification (κ coefficient = 1). Taken together, a real-time RPA method for APV screening was established in this study.
