The standardization of One-step real time reverse transcription PCR assay is a specific, sensitive, rapid and cost-effective assay for detection influenza A virus stains i.e. H1N1, H3N2 are responsible for the seasonal influenza infection. Influenza A virus can be diagnosis by the variety of diagnostic tools like isolation of virus, conventional reverse transcription PCR and real time reverse transcription PCR assays. Although conventional reverse transcription PCR is specific, sensitive, rapid and cost effective also but there is need of post reverse transcription PCR analysis. However conventional reverse transcription PCR only gives the qualitative result not the quantitative results but in case of real time reverse transcription PCR hence less time consuming and less chance of cross contamination as compared with the conventional reverse transcription PCR. There is no need of post reverse transcription PCR analysis are required and gives the qualitative as well as quantitative result with less chance of cross contamination. Only the negative point of real time reverse transcription PCR is high cost instrument is required for the assay.
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