LAMP assay is an auxiliary tool for rapid and sensitive pathogen diagnosis in veterinary areas

Kittisak Buddhachat

doi:10.12982/vis.2022.019

Abstract

Molecular technique for nucleic acid amplification has become more advanced, especially isothermal nucleic acid amplification which can amplify DNA or RNA molecules at constant temperatures. This allows us to be independent of sophisticated equipment such as thermal cyclers or realtime-PCR. The feasibility of adopting molecular techniques for pathogen detection at point-of-care are approaching practical use. At present, there are various isothermal nucleic acid amplification technologies, including rolling cycle amplification (RCA), helicase-dependent amplification (HDA), recombinase polymerase amplification (RPA) and loop-mediated isothermal amplification (LAMP). LAMP is one of the most popular methods used for pathogen diagnosis in animals due to several readout platforms such as turbidity, fluorescence, lateral flow dipstick (LFD) and color change visualizable by naked eye. In addition, LAMP can be integrated with CRISPR-cas which in turn, augments pathogen detection at ultra sensitivity. This letter shows the implementation of LAMP for pathogen detection in veterinary areas with different readouts as well as a new perspective for LAMP adaptation.

Highlights

Molecular technique for nucleic acid amplification has become more advanced, especially isothermal nucleic acid amplification which can amplify DNA or RNA molecules at constant temperatures
Isothermal nucleic acid amplification has been established in various methods such as rolling cycle amplification (RCA), helicase-dependent amplification (HDA), recombinase polymerase amplification (RPA) and loop-mediated isothermal amplification (LAMP) (Craw et al, 2015)
DNA-binding dyes that act as intercalating dyes such as SYBR green I, SYTO9, Gel®Red can be added at the endpoint of the LAMP solution to observe the result by color change under the naked eye or fluorescence (Quyen et al, 2019)

Summary

Introduction

Molecular technique for nucleic acid amplification has become more advanced, especially isothermal nucleic acid amplification which can amplify DNA or RNA molecules at constant temperatures. Many platforms of molecular testing (e.g., PCR-based assay, isothermal nucleic acid amplification, and CRISPR-cas-based assay) are promptly implemented for rapid diagnostic test (Böger et al, 2021).

Results

Conclusion