Submit or Track your Manuscript LOG-IN

Comparative Detection of 1-15N1 Avian Influenza Virus using Conventional RT- PCR and Real Time RT-PCR

Comparative Detection of 1-15N1 Avian Influenza Virus using Conventional RT- PCR and Real Time RT-PCR

Hagag, N.M.*•, Arafa, A.*; Shalaby, M. A. ** El-Sanousi, A. A. **and Aly, M. M. 

*National Laboratory for Veterinary Quality Control on Poultry Production, P.O. Box 264, Dokki, Giza.
** Department of Virology, Faculty of Veterinary Medicine, Cairo University12211Giza-Egypt

ABSTRACT

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 AlVs in biological specimens is critical for limiting further spread of the disease in poultry. Application of molecular methods such as Reverse Transcriptase polymerase chain reaction (RT-PCR) and Real time RT-PCR (RRT-PCR) as a rapid, specific and sensitive detection methods currently used in national and reference laboratories worldwide. In this study a comparison of the specificity and sensitivity between 2 different formats of conventional RT-PCR (one and two steps) and 3 different formats of RRTPCR (one step using TaqMan probe, two steps using TaqMan probe and two steps using hybridization probe) were performed and compared as a diagnostic tool for H5N1 virus detection. All these formats of PCRs appeared within the same specificity for H5 gene detection, while they showed difference in sensitivity as the one step conventional RT-PCR showed to be more sensitive than two steps conventional RT-PCR by 10 folds, one step RRT-PCR TaqMan probe was more sensitive than two steps RRT-PCR TaqMan probe by 10 folds, two steps RRT-PCR hybridization probe is more sensitive than two steps RRT-PCR TaqMan probe by 100 folds, finally two step RRT-PCR using hyperidization probe is more sensitive than conventional RT-PCR two steps by 1000 folds. 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.

To share on other social networks, click on any share button. What are these?

Journal of Virological Sciences

July

Vol. 3, Iss. 1

Featuring

Click here for more

Subscribe Today

Receive free updates on new articles, opportunities and benefits


Subscribe Unsubscribe