Development of a loop-mediated isothermal amplification (LAMP) assay for the rapid detection of Beiji nairovirus

Beiji nairovirus (BJNV) is a recently discovered tick-borne RNA virus associated with human febrile illness. This study aimed to develop a loop-mediated isothermal amplification (LAMP) assay for the precise detection of BJNV, with a specific focus on assessing its effectiveness with clinical samples. The optimal molecular target was identified as the BJNV small (S) segment gene, and the ideal reaction conditions were established at 65 °C for 50 min. A neutral red stain concentration of 300 μM was determined to be optimal for visualizing the LAMP reaction. The LAMP method demonstrated an impressive lower limit of detection at 10 copies/μL, highlighting a sensitivity level 10,000 times higher than traditional PCR methods. Moreover, it showed no cross-reactivity with three other viruses. The LAMP assay demonstrated results consistent with those of semi-nested PCR when applied to clinical samples, thereby validating its suitability for field testing. In conclusion, the LAMP assay developed in this study represents a significant advancement in the rapid detection of BJNV. Its high sensitivity, specificity, and ease of use make it a valuable tool for the establishment of effective prevention and control strategies against this emerging pathogen.