Rapid diagnosis of infectious Laryngo tracheitis virus (ILTV) by using molecular method of real-time PCR, based on UL27 proprietary gene

Infectious Laryngo tracheitis virus (ILTV) is one of the major pathogens in poultry industry, economically. Poultries are the only reservoir of the virus. This agent causes a relatively prevalent and acute respiratory syndrome in laying Hens and in particular in broiler breeder hens, which reduces egg production and increases mortality in them. Since at present, unfortunately, it does not exist a rapid, sensitive and accurate method for detection of the virus yet, so in this research our aim is to evaluate and introduce an accurate molecular method by using real-time PCR technique, based on UL27 proprietary gene of ILTV, in order to diagnose the virus, rapidly. In this study, after preparation of viral sample, respective DNA was extracted from virus by using viral nucleic acids extraction kit. Then, the existence of UL27 proprietary gene of the virus was evaluated at first by traditional PCR technique using specific primers and afterwards by real-time PCR technique using specific probe and primers. Obsvervation of expected 96 bp fragment in electrophoresis gel analysis, confirmed the presence of UL27 proprietary gene in this viral samples and the results of real-time PCR assay evaluation were also positive in this samples. This study showed that the molecular method of real-time PCR is the proper method for rapid and accurate diagnosis of ILTV by detection of UL27 proprietary gene.