Two in-House One-Step rRT-PCR Assays, Developed for Accurate and Rapid Molecular Identification of Newcastle Disease Virus, on the basis of SYBR Green and Specific TaqMan Probe

Background and Aims: Newcastle disease virus (NDV) is an avian paramyxovirus (A-PMV 1) and one of the major pathogens in poultries. Vaccination is intended to control the disease, nevertheless this virus is a growing threat to the poultry industry. So, early detection of the virus can prevent the spread of illness and avoid huge economic losses. Towards this goal, in this research, we developed two novel in-house one-step rRT-PCR assays based on SYBR Green and specific TaqMan probe for accurate and rapid molecular identification of Newcastle disease virus. Materials and Methods: In this experimental study, after preparation of viral sample, respective RNA was extracted from virus by using viral RNA extraction kit. The specific probe and primers were designed, based on the conserved region of matrix (M) protein encoding gene of NDV, and used to produce c-DNA and subsequently to amplify and detect this region by two novel one-step real-time reverse-transcription polymerase chain reaction (rRT-PCR) assays. These diagnostic appraisements were carried out using SYBR Green and matrix (M) gene specific TaqMan probe. Results: The presence of M gene in RNAs, extracted from NDV samples, was confirmed by both of these one-step rRT-PCR assays and the results of these diagnostic tests were positive. Conclusion: This study showed that these two developed one-step rRT-PCR assays are the proper molecular methods for rapid and accurate diagnosis of NDV by detection of matrix (M) protein encoding gene.