Molecularybiological diagnostics of respiratory diseases in birds

Abstract

Respiratory diseases of birds cause significant economic loses to poultry farming. In the analysis of these diseases, a significant role belongs to metapneumovirus infection in birds and infectious bronchitis in chickens. These diseases often occur in complex with other viral and bacterial infections. Isolation of these viruses from the test material using known methods is a rather difficult and time-consuming process. The difficulty in isolation exists cause of the very short stage of accumulation of viruses in significant titers in the organs of birds. Therefore, nowadays, the main role in the correct diagnosis of these diseases belongs to molecular biological methods, in particular, polymerase chain reaction (PCR) and sequencing. The main objective of the research was to develop universal primers that allow the identification of pathogens of avian and IBC chickens, and serotyping of these viruses by electrophoretic detection and sequencing based on PCR. As a result of assays it was established that - presence of PCR products of the cDNA fragment with a length of 291 BP indicates the presence in the material RNA of SSV subtype, and the cDNA fragment with a length of 501 BP - for the RNA of SSV subtype A. the presence in the sample of the virus genome BC is confirmed by the detection of a fragment with a length of 526 BP. Obtained sequenced primers showed high specificity in relation to the diagnosed pathogens, which allows them to be successfully used in research institutions conducting work on the study of these viruses and molecular diagnosis of the corresponding infections.