Loop-mediated isothermal amplification combined with lateral flow biosensor for rapid and sensitive detection of monkeypox virus

Front Public Health. 2023 Mar 24:11:1132896. doi: 10.3389/fpubh.2023.1132896. eCollection 2023.

Abstract

The ongoing outbreak of the monkeypox, caused by monkeypox virus (MPXV), has been a public health emergency of international concern, indicating an urgent need for rapid and sensitive MPXV detection. Here, we designed a diagnostic test based on loop-mediated isothermal amplification (LAMP) and nanoparticle-based lateral flow biosensor(LFB)for diagnosis of MPXV infection, termed MPX-LAMP-LFB. A set of six LAMP primers was designed based the ATI gene of MPXV, and LAMP amplification of MPXV templates was performed at 63°C for only 40 min. The results were rapidly and visually decided using the LFB test within 2 min. The MPX-LAMP-LFB assay can specifically detect MPXV strains without cross-reaction with non-MPXV pathogens. The sensitivity of the MPX-LAMP-LFB assay is as low as 5 copies/μl of plasmid template and 12.5 copies/μl of pseudovirus in human blood samples. The whole process of the MPX-LAMP-LFB assay could be completed ~1 h, including rapid template preparation (15 min), LAMP reaction (40 min)and result reporting (<2 min). Collectively, MPX-LAMP-LFB assay developed here is a useful tool for rapid and reliable diagnosis of MPXV infection.

Keywords: diagnosis; lateral flow biosensor; loop-mediated isothermal amplification; monkeypox virus; nanoparticles.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biosensing Techniques* / methods
  • Humans
  • Monkeypox virus* / genetics
  • Monkeypox virus* / isolation & purification
  • Mpox (monkeypox) / diagnosis
  • Nucleic Acid Amplification Techniques
  • Sensitivity and Specificity

Supplementary concepts

  • LAMP assay