Comparison of Reverse Transcription Loop-Mediated Isothermal Amplification Method with SYBR Green Real-Time RT-PCR and Direct Fluorescent Antibody Test for Diagnosis of Rabies

Rabies as an endemic disease in most Asian and African countries, especially in remote areas, and requires a reliable diagnostic method. This study aimed to develop a reverse transcription loop-mediated isothermal amplification (RT-LAMP) method for rapid detection of rabies virus RNA in the brain samples, compared to SYBR Green real time RT-PCR test as a molecular technique and direct fluorescent antibody test as a serological method. In this study, RT-LAMP was developed to diagnose rabies. Six primers were designed based on the nucleoprotein (N) of rabies virus. The sensitivity and specificity of SYBR Green real-time RT-PCR and RT-LAMP methods were also determined.RT-LAMP was optimized at 58 ℃ for 60 min. The sensitivity and specificity of RT-LAMP and SYBR Green real-time RT-PCR were 91.2% and 84.2%, and 94.12% and 88.9%, respectively. The slight difference between the sensitivity and specificity of RT-LAMP and that of SYBR Green Real-Time RT-PCR demonstrated that RT-LAMP could be used as a reliable and cost-effective method for the diagnosis of rabies.