Development of a HiFi-LAMP Assay for Detection of Human Pegivirus Type 1 RNA

J Med Virol. 2025 Jun;97(6):e70423. doi: 10.1002/jmv.70423.

Abstract

Human pegivirus (HPgV) is a blood-borne RNA virus belonging to the family Flaviviridae, and contains two types HPgV-1 and HPgV-2. HPgV-1 infection does not cause acute diseases in healthy individuals but was demonstrated to play beneficial roles in individuals coinfected with HIV-1. HPgV-1 has a high prevalence in blood donors and the general population worldwide. The long-term consequence of HPgV-1 infection in healthy individuals remains unknown. High prevalence of HPgV-1 in blood donors raises concerns about the risk of transfusion transmission. Development of rapid and accurate point-of-care testing (POCT) of HPgV-1 will facilitate 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 limits of detection (LODs) of 122.6-135.7 copies of viral RNA/25 μL reaction for various HPgV-1 variants and can be completed within 30 min. 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.

Keywords: HiFi‐LAMP; Human pegivirus type 1 (HPgV‐1); RT‐qPCR; healthy adult; transfusion; untranslated region (UTR).

Publication types

  • Evaluation Study

MeSH terms

  • Adult
  • Blood Donors
  • China / epidemiology
  • Female
  • Flaviviridae Infections* / diagnosis
  • Flaviviridae Infections* / epidemiology
  • Flaviviridae Infections* / virology
  • Humans
  • Limit of Detection
  • Male
  • Middle Aged
  • Molecular Diagnostic Techniques* / methods
  • Nucleic Acid Amplification Techniques* / methods
  • Pegivirus* / genetics
  • Pegivirus* / isolation & purification
  • Point-of-Care Testing
  • RNA, Viral* / genetics
  • Sensitivity and Specificity
  • Young Adult

Substances

  • RNA, Viral

Supplementary concepts

  • LAMP assay