Rapid detection of classical swine fever virus by loop-mediatedisothermal amplification combined with lateral flow dipstick method

Objective The experiment was conducted to establish a rapid real time detection method of classical swine fever virus at local fields.

Method A rapid detection method(RT-LAMP-LFD) of classical swine fever virus(CSFV) was developed by loop-mediated isothermal amplification (LAMP) combined with lateral flow dipstick (LFD) method. The LAMP method for amplification of the CSFV NS5B gene using fluorescein isothiocyanate-labeled primer was combined with a chromatographic LFD for rapid and simple visual detection of CSFV.

Result This method could specifically detect CSFV, but not other virus such as PRRSV, JEV and PCV-2. The minimum detection limit of the CSFV-RT-LAMP-LFD method was 50 pg. The detection could be finished within 1 h without using complex instruments such as PCR machine and the results were visible by eye.

Conclusion This RT-LAMP-LFD method is a highly specific and sensitive, safe, rapid and simple way for field detection of CSFV infection.