Evaluation and optimization of loop-mediated isothermal amplification (LAMP) technique in capripoxvirus diagnosis and its comparison with PCR method

Sheep pox (SP), goat pox (GP), and lumpy skin disease (LSD), subspecies of the Capripox virus (CaPVs), are important in the pathogenesis of sheep, goats, and cattle. The causative agent is the capripox virus which has been isolated in South Africa for the first time. Sheeppox (SP), goatpox (GP), and lumpy skin disease (LSD) viruses are morphologically indistinguishable from each other and have been adapted to different host species (۴). From a serological point of view, it is difficult to differentiate these viruses and cross-immunity occurs among them (۲). The present study reports the evaluation and optimization of a novel loop-mediated isothermal amplification (LAMP) technique for the rapid detection of CaPVs and compares the LAMP technique with the PCR method. LAMP primers were selected from the P۳۲-protected gene of the Capripox virus. Safe-Red fluorescent dye was used to monitor and change the color in the disease’s positive cases to bright yellow at a ۳۲۰-nm wavelength, and the final results were confirmed using electrophoresis. The proposed LAMP test for the Capripox virus showed high specificity and non-cross-reactivity with viruses of Poxviridae with similar clinical symptoms. The optimized LAMP test was compared with PCR. LAMP and PCR were similar in diagnostic sensitivity. The specificity was evaluated using ۳۰ samples of cow skin suspected of having lumpy skin disease as well as ۱۶ samples of positive and negative references and the negative control. The proposed easy-to-use, inexpensive, high-sensitivity LAMP test might be suitable for detecting caprypox virus in laboratories in border areas and rural areas.

کلیدواژه ها:

LSDV ، LAMP ، PCR ، Capripoxvirus