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Comparative reproducibility of SYBR Green I and TaqMan real-time PCR chemistries for the analysis of matrix and hemagglutinin genes of Influenza A viruses

Background: The outbreak of novel influenza A H1N1-2009 virus (pH1N1) and its rapid spread worldwide raised serious concern about pandemic preparedness.

Objectives: The present study was designed to evaluate the sensitivity and specificity of two chemistries of real-time RT-PCR (with the use of fluorescent SYBR Green I dye and specific TaqMan probe) for detection of the matrix and hemagglutinin genes of human influenza A viruses.

Methods: Influenza A virus reference strains were used to perform the calibration curve analysis and the diagnostic accuracy of both the real-time RT-PCR formats were assessed on 110 clinical specimens from patients presenting with influenza-like-illness (ILI).

Result: The study results suggest that the TaqMan chemistry has better sensitivity as compared to SYBR Green I, however, the SYBR Green I assay was found to be simpler, economical and readily available. Conclusions: It is concluded that the SYBR Green I assay has equal potential to be considered as an alternative to the TaqMan assay as a preparedness measure for any future influenza outbreak management.

Author(s): Binod Kumar, Prashant Kumar, Roopali Rajput , M K Daga, Varinder Singh , Madhu Khanna

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