Comparative Detection of H5N1 Avian Influenza Virus Using Conventional Rt-PCR and Real Time Rt-PCR

• Published in: International journal of virology Vol. 8; no. 2; pp. 203 - 213
• Main Authors: Hagag, N.M., Arafa, A., Shalaby, M.A., El-Sanousi, A.A., Aly, M.M.
• Format: Journal Article
• Language: English
• Published: 2012
• Subjects: Avian influenza virus; Egypt, Arab Rep
• ISSN: 1816-4900
• DOI: 10.3923/ijv.2012.203.213

Highly Pathogenic Avian Influenza (HPAI) caused by influenza A H5N1 virus, poses a significant threat to the poultry industry and humans in Egypt. Since it was first recognized in 2006, the disease has become enzootic in poultry throughout Egypt and still circulates in the poultry population, so the ability to rapidly recognize AIVs in biological specimens is critical for limiting further spread of the disease in poultry. In this study a comparison of the specificity and sensitivity between 2 different formats of conventional RTPCR (one and two steps) and 3 different formats of RRTPCR (one step using TaqMan(R) probe, two steps using TaqMan(R) probe and two steps using hybridization(R) probe) were performed and compared as a diagnostic tools for H5N1 virus detection. Fifty one field samples were further tested by all mentioned PCR formats the results were the same of that obtained in the sensitivity experiment and agreed with them in placing hybridization probe system of higher sensitivity than TaqMan one step than TaqMan two steps and conventional PCR were of the lowest sensitivity although one step showed higher sensitivity than two steps.