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Published on: August 2, 2011
Development of a new TaqMan real-time PCR for detecting Psittacine adenovirus 1 (PsAdV-1)
Laura Andrea Charola-Ramos1, Nira Vega-Pita2, María José Bernal-Guadarrama3
1Instituto Universitario de Enfermedades Tropicales y Salud Pública de Canarias, Universidad de La Laguna, Avda. Astrofísico Fco. Sánchez, S/N, San Cristóbal de La Laguna, 38203, Spain. lcharola@ull.edu.es.
Background:
Psittacine adenoviruses belong to the Adenoviridae family, that includes genera Aviadenovirus, Barthadenovirus, and Siadenovirus and may cause either fatal disease or subclinical infections. Subclinical infections are particularly relevant when infected birds enter the bird trade, as infected birds can act as sources of viral spread. Given the ease of transmission of these viruses, the availability of sensitive and specific diagnostic tools is essential. The purpose of this study was to establish a TaqMan-based quantitative PCR (qPCR) assay for the detection and quantification of Psittacine adenovirus 1 (PsAdV-1) in psittacine samples.
Results:
Serial dilutions of a control plasmid and clinical samples were analysed using conventional PCR, SYBR Green qPCR and the newly developed TaqMan qPCR. A psittacine-specific 18 S rRNA TaqMan assay was used as an endogenous control to assess DNA extraction quality and sample suitability, particularly in cloacal swabs. All three methods showed comparable sensitivity when applied to liver samples. However, the TaqMan assay demonstrated improved detection in cloacal swabs compared to conventional PCR, while maintaining high specificity and showing no cross-reactivity with other avian pathogens.
Conclusions:
The TaqMan-based qPCR assay developed in this study provides a rapid, sensitive and highly specific method for the detection of PsAdV-1 in clinical samples. Its improved performance in cloacal swabs makes it a valuable tool for routine diagnosis and epidemiological surveillance of psittacine adenovirus infections, particularly in asymptomatic birds.
