Efficiency of Three Commercial Real-Time PCR Kits for Detecting Classical Swine Fever Virus

Abstract

A comparative study of the efficiency of three commercial real-time PCR kits for detecting Classical Swine Fever Virus (CSFV) by real-time RT-PCR from 10-fold serially diluted viral plasmid DNA samples was conducted. The experiment was conducted three times, each trial using a different commercial kit but maintaining the same concentrations of probes and primers. The template volume and the number of real-time RT-PCR cycles (45 cycles) were also kept constant. The results showed that the TaqMan^® Fast Virus 1-Step Master Mix kit was able to detect diluted plasmid DNA of classical swine fever virus at the lowest concentration of 7.923 copies/µL. Furthermore, it achieved detection with a lower cycle number compared to the SuperScript™ III Platinum™ One-Step qRT-PCR Kit and QuantiTect® Probe RT-PCR Kit at each corresponding concentration. The PCR efficiency (E) and $R^2$ values for the TaqMan^® Fast Virus 1-Step Master Mix were also within the acceptable range. Therefore, the TaqMan^® Fast Virus 1-Step Master Mix kit was selected for the detection of classical swine fever virus by real-time RT-PCR using the QuantStudio™ 5 Real-Time PCR System.