Rapid detection of Influenza A (H1N1)Viruses by RT-PCR & sequencing methods

Negative sence RNA genome of Influenza A virus contains 8 segments coding for 12-14 proteins depending on strains. Genetically modified virus is caused world wide spread of a new Influenza in the human population. Developing a rapid and accurate diagnostic method to identify new species is necessary. The aim of this study was rapid detection of new species of Influenza A subtypes using specific RT-PCR based on hemagglutinin gene.
In this study 30 Nasopharynx samples of patient cultured in embryonated eggs. Then RNA was extracted, cDNA prepared and PCR was performed using specific primers designed from hemagglutinin gene. PCR products purified and sequenced.
Findings: PCR products sequences compared with Influenza A sequences obtained from the Gene Bank database. All positive isolates most closely related to the influenza reference strain. This Result showed that the specific RT-PCR used was able to amplified and detect Influenza A subtypes from clinical specimen.
The results of this study confirmed that PCR based on hemagglutinin gene with sequencing is a sensitive and accurate method for rapid detection of influenza A new subtypes directly from clinical specimen which is useful in preparation and production of vaccine.