Real-time reverse transcription PCR for detection and quantitative analysis of equine influenza virus

Abstract

Equine influenza is a cause of epizootic respiratory disease of the equine. The detection of equine influenza virus using real-time Light Cycler reverse transcription (RT)-PCR technology was evaluated over two influenza seasons with the analysis of 171 samples submitted for viral respiratory disease. Increased sensitivity was found in overall viral detection with this system compared to Directigen Flu A and virus isolation, which were 40% and 23%, respectively, that of the RT-PCR. The assay was also evaluated as a viable replacement for the more traditional methods of quantifying equine influenza virus, 50% egg infectious dose and 50% tissue culture infectious dose. There was a significant positive correlation (P<0.05) between the quantitative RT-PCR and both of these assays.

FIG. 1.

Mean and standard error of the mean of viral RNA copies/ml (log₁₀) in nasal secretions of four foals from days 2 to 7 postchallenge with equine influenza virus.