Development of real-time PCR assays for single and simultaneous detection of infectious bursal disease virus and chicken anemia virus

Publication date: Available online 14 November 2018Source: Molecular and Cellular ProbesAuthor(s): Claudia Techera, Gonzalo Tomás, Yanina Panzera, Alejandro Banda, Paula Perbolianachis, Ruben Pérez, Ana MarandinoAbstractInfectious bursal disease virus (IBDV) and chicken anemia virus (CAV) cause relevant immunosuppressive diseases in poultry. Clinical diagnosis of these viruses is challenging given the different disease presentations and the frequent occurrence of co-infections with other pathogens. Here, we standardized and validated simplex and duplex RT-qPCR assays for the straightforward detection of IBDV and CAV. The qPCR assays are based on primers and hydrolysis probes that target highly conserved regions of IBDV and CAV genomes. Analytical sensitivity tests on 10-fold serial dilutions containing 100-108 viral genomes indicated that the simplex assays have good determination coefficients and efficiency and detect a wide range of virus doses (102 to 108 molecules copies/reactions). The relatively small values of intra- and inter-assay variability ensure the repeatability and support its reproducibility in different diagnostic and research facilities. The assays are also efficient tools for absolute quantification as indicated by the analytical performance analysis. The assays have an excellent specificity and absence of cross-reactivity with negative samples, or with other common avian viruses. The simplex IBDV and CAV assays use probes labelled with different dyes (FA...
Source: Molecular and Cellular Probes - Category: Molecular Biology Source Type: research