Detalhes do Artigo
[305909] Comparison of RT-qPCR and RT-ddPCR methodologies for the detection of Bovine Viral Diarrhea Virus (BVDV) RNA
Autores: Ariane de Mesquita Rodrigues, Aurea Valadares Folgueras Flatschart, Roberto Becht Flatschart, Mayane Ribeiro de Faria Henrique, José Lafaiete Palles Ramos Júnior
Resumo: The Bovine Viral Diarrhea Virus is responsible for causing infection of great importance in cattle, being highly disseminated in the world and endemic in Brazil. This virus is also described as the main contaminating virus of Fetal Bovine Serum (FBS), a supplement routinely used in cell cultures and which can become a source of contamination for these cells. Qualitative methods for detecting BVDV RNA can be used both for the diagnosis of the disease in animals and to detect the infection of cultures. In this study, we compared the results of a commercial diagnostic method by Real Time RT-PCR (RT-qPCR) to the RT-Droplet Digital PCR (RT-ddPCR) method using the same set of probe and primers. Serial dilutions of the BVDV RNA standard from the commercial kit and also the BVDV RNA produced in MDBK cells were evaluated by both methods. RT-qPCR showed 100% sensitivity and 80% to 83% specificity compared to the RT-ddPCR method. Points of disagreement between the two techniques arose at low RNA concentrations.