Detection of human pegivirus type 1 RNA in healthy adults by a novel HiFi-LAMP assay

Human pegivirus (HPgV) is a blood-borne RNA virus belonging to the family Flaviviridae , and contains two types HPgV-1 and HPgV-2. As a non-pathogenic human virus, HPgV-1 has a high prevalence in blood donors and general population worldwide, and was demonstrated to play beneficial roles in individuals co-infected with HIV-1. High prevalence of HPgV-1 in blood donors raises concerns about the risk of transfusion transmission. Rapid and accurate point-of-care testing (POCT) of HPgV-1 facilitates the screening of HPgV-1 infection among blood donors. Here, we reported a novel high-fidelity loop-mediated isothermal amplification (HiFi-LAMP) assay for detection of HPgV-1 and evaluated its performance in 175 healthy adults from Taizhou, China. The assay exhibits high specificity and sensitivity with a limit of detection (LOD) of 148 copies/25 μL reaction and can be completed within 30 minutes. Clinical validation showed that the assay had a 100% concordance with a previously described RT-qPCR assay for 175 sera from healthy adults, showing 100% sensitivity and specificity. Furthermore, we reported a 28.0% (111/397) prevalence of HPgV-1 in healthy adults in Taizhou, China, with no significant differences between genders and ages. The prevalence is obviously higher than a pooled HPgV-1 prevalence of 3.3% in blood donors in China. The novel HPgV-1 HiFi-LAMP assay offers a robust, rapid, and cost-effective tool for HPgV-1 surveillance to mitigate transfusion risk especially in resource-limited areas. High prevalence of HPgV-1 in healthy adults underscores its potential public health relevance.