Direct Detection of SARS-CoV-2 RNA in Saliva with Colorimetric RT-LAMP

Methods Mol Biol. 2024:2822:39-50. doi: 10.1007/978-1-0716-3918-4_4.

Abstract

RT-LAMP is an effective alternative to RT-PCR-based diagnostics, offering high specificity, sensitivity, and rapid results. One notable advantage is the robustness of its enzymes, allowing for direct amplification from crude samples without the need for prior isolation of RNA. Colorimetric LAMP is particularly attractive as it eliminates the need for complex instrumentation, making it suitable for point-of-care applications. Here, we present a comprehensive step-by-step protocol for establishing an RT-LAMP-based test for direct detection of SARS-CoV-2 genomic RNA in saliva samples using different colorimetric detection methods. Importantly, this versatile test can be easily adapted to detect emerging pathogens.

Keywords: Br-PAPS; Colorimetric; Diagnostic; HNB; LAMP; Murexide; Phenol red; RNA virus; Saliva.

MeSH terms

  • COVID-19 Nucleic Acid Testing / methods
  • COVID-19* / diagnosis
  • COVID-19* / virology
  • Colorimetry* / methods
  • Humans
  • Molecular Diagnostic Techniques* / methods
  • Nucleic Acid Amplification Techniques* / methods
  • RNA, Viral* / analysis
  • RNA, Viral* / genetics
  • RNA, Viral* / isolation & purification
  • SARS-CoV-2* / genetics
  • SARS-CoV-2* / isolation & purification
  • Saliva* / virology
  • Sensitivity and Specificity

Substances

  • RNA, Viral

Supplementary concepts

  • LAMP assay