Droplet Digital Polymerase Chain Reaction Method for Absolute Quantification of Influenza A Viruses.

Abstract

Digital polymerase chain reaction (dPCR) is a new method for absolute quantification. However, an absolute quantification method for influenza A viruses based on droplet digital polymerase chain reaction (dPCR) has not been established. In this study, we found that: (i) the annealing temperature of dPCR was optimized at 64, 4°C; (ii) the detection range of dPCR was 37.7 approximately 8.22 X 10(4) copies/μl for influenza A viruses; (iii) the limit of detection of dPCR was 3. 77 copies/reaction. The liner correlation coefficient (R(2)) was found to be 0. 9988, suggesting the high reliability of our dPCR method. The dPCR method could be used to detect influenza A viruses clinically. In summary, we developed a dPCR method for absolute quantification of influenza A viruses. This method could be used effectively to quantify influenza A viruses in clinical samples. Therefore, our method could be a new tool for the absolute quantification of viral load.