Development of a Molecular Genetic Diagnostic Kit for the Detection of the Avian Influenza Virus H5N1

Introduction

Introduction of an influenza A virus with a novel HA gene in a population which lacks immunity to this HA has the potential to cause a pandemic when the virus possesses the ability to spread efficiently among humans. In order to limit the spread of highly pathogenic avian influenza(HPAI), rapid detection and identification of the infectious agent are required. The current pandemic spread of the H5N1 viruses in Egypt as well as many other countries urged the need to develop a proper strategy for the control of this infectious agent. The first step of this strategic plan is to develop a reliable, specific, accurate and cost effective protocols for the identification of the virus. The current situation of H5N1 in Egypt also necessitate the development of diagnostic materials in order to facilitate the detection of H5N1.

Objectives

The specific objective of this proposal was to develop a cost effective,  sensitive, and accurate molecular genetic diagnostic kit for the detection of the Avian Influenza H5N1 viruses using single step, single tube Multiplex RT-PCR .

Duration:                 One Year

Starting date:          March, 2006

Results

- The first diagnostic kit for the molecular genetic diagnosis of the H5N1 viruses using a single tube single step RT-Multiplex PCR protocol was developed . Patency by the Egyptian Academy for Science and technology was applied for in August, 2006. (Patent file No. 206080398).

- The second diagnostic kit was developed in December, 2006. In this molecular genetic diagnostic kit the M1 gene primer set was added to  the H5N1 primers in order to detect the M1 gene in addition to the H5N1 genes. This addition gives us the ability to monitor the occurrence of new H5N1 mutant strains in a single tube, single step RT-Multiplex PCR protocol. Filing of a new patent is now in progress.