Design, Optimization, and Analytical Performance Evaluation of LAMP-Based Rapid Detection Assay for Severe Fever with Thrombocytopenia Syndrome Virus

Abstract

Objectives: Severe Fever with Thrombocytopenia Syndrome Virus (SFTSV), a tick-borne pathogen, presents a growing public health threat in East Asia. Although conventional RT-PCR methods are effective for detection, they are limited by the need for specialized equipment and time-consuming procedures. This study aimed to develop and evaluate a rapid, sensitive, and field-deployable diagnostic method for SFTSV using Loop-Mediated Isothermal Amplification (LAMP). Methods: Twelve sets of LAMP primers were designed to target the RNA-dependent RNA polymerase (L segment) of the SFTSV genome. These primers were screened through stepwise colorimetric LAMP assays to identify the optimal set. The sensitivity and specificity of the selected primer set were evaluated using serial dilutions of SFTSV RNA and a panel of control pathogens. Results: The selected LAMP primer set demonstrated high amplification efficiency, successfully detecting as little as 10 attograms (ag) of SFTSV RNA. Moreover, the primer set showed no cross-reactivity with non-SFTSV samples, confirming its high specificity. Conclusion: The developed LAMP assay provides a rapid and reliable method for SFTSV detection, with potential for use in field settings. This diagnostic tool could enhance early detection and improve outbreak response for SFTSV infections.