Performance of One Tube RT-PCR Assay in Rapid Detection of Influenza Virus
Authors: Broomand. Chacharaein, Aini Ideris, Abdul Rahman Omar and Khatijah Yusoff
Faculty of Veterinary Medicine, Faculty of Biotechnology and Biomolecule Sciences, Universiti Putra Malaysia 43400 UPM Serdang Selangor Malaysia.
Avian influenza is a viral disease affecting the respiratory, digestive and/or nervous systems of many species of birds, both domestic and wild. Universiti Putra Malaysia (UPM) researchers developed a one tube NP RT-PCR which can quickly identify the bird flu virus.
The NP RT-PCR was evaluated using 20 different influenza A strains and showed that it is able to amplify the nucleoprotein (NP) gene from a series of different avian influenza. In other words, it was able to identify a series of different bird flu viruses.
The method was also able to amplify the NP and HA gene extracted from cloacal, tracheal and organ samples, which were collected from experimentally infected SPF (specific pathogen free) chickens. The ability of influenza A virus detection by the NP RT_PCR assay, was estimated to be as sensitive as virus isolation by inoculation in SPF eggs.
Further tests were made to determine how specific the technique was for identifying influenza A viruses. This was done by including RNA from Newcastle disease virus, infectious bronchitis virus and infectious bursal disease virus. No amplification was observed for any of these viruses thus further confirming that the NP RT-PCR technique is specific to influenza A viruses only.
A general influenza diagnostic RT_PCR is not only important for the detection of virus from an actual outbreak but also for the screening of potential carriers of influenza A. In conclusion, the NP RT-PCR is a rapid and sensitive method for the detection of influenza virus from clinical samples.
