Rapid diagnosis of infectious Laryngo tracheitis virus (ILTV) by real-time PCR on UL27 proprietary gene

Subject Areas : Virology

Samaneh Zahiriyeganeh ¹ , Mohammad Sadegh Hashemzadeh ² , Ehsan Zafari ³ , Mahdi Tat ⁴ , Mohammad Najarasl ⁵ , Bentolhoda Zahraei ⁶ , Ruhollah Dorostkar ⁷

1 - M.Sc., Applied Virology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran.
2 - Ph.D student, Applied Virology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran.
3 - Ph.D student, Applied Virology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran.
4 - M.Sc., Applied Virology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran.
5 - Ph.D student, Applied Virology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran.
6 - M.Sc., Applied Virology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran.
7 - Assistant Professor, Applied Virology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran.

Received: 2015-06-05 Accepted : 2015-07-29 Published : 2015-12-01

Keywords: Real-time PCR, Infectious Laryngo tracheitis virus (ILTV), UL27 gene,

Abstract :

Background & Objectives: Infectious Laryngo tracheitis virus (ILTV) is one of the major pathogens causing economic losses in poultry industry. Poultries are the only reservoir of the virus. By now there is no rapid, sensitive and accurate method for detection of this virus. This study was aimed to evaluate and introduce an accurate molecular method by using real-time PCR technique on UL27 proprietary gene of ILTV in order to fast diagnosis of the virus. Materials & Methods: In this experimental study, after preparation of viral sample, the DNA was extracted from virus by using viral nucleic acids extraction kit. Thereafter, 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. Results: Observation 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. Conclusion: This study showed that the molecular method of real-time PCR is a suitable method for rapid and accurate diagnosis of  ILTV by detection of UL27 proprietary gene.