A loop-mediated isothermal amplification (LAMP) assay for rapid detection of Anaplasma phagocytophilum infection in dogs

Abstract

A simple and rapid diagnostic method of Anaplasma phagocytophilum infection was established, employing the loop-mediated isothermal amplification (LAMP) reaction. With 1 set of 4 specific primers targeting the citrate synthase (gltA) gene of A. phagocytophilum, the LAMP assay rapidly amplified the target DNA sequence in a single tube in 1 h, requiring only a water bath for reaction. When visualized by gel electrophoresis, the LAMP products appeared as a ladder-like pattern, with many bands of different sizes from 180 base pairs up to the loading well. The results obtained from testing 32 clinical blood samples of dogs demonstrated that the LAMP method was more sensitive than nested PCR in the diagnosis reaction for the detection of A. phagocytophilum DNA.