Abstract

The rampant spread of COVID-19 and the worldwide prevalence of infected cases demand a rapid, simple, and cost-effective Point of Care Test (PoCT) for the accurate diagnosis of this pandemic. The most common molecular tests approved by regulatory bodies across the world for COVID-19 diagnosis are based on Polymerase Chain Reaction (PCR). While PCR-based tests are highly sensitive, specific, and remarkably reliable, they have many limitations ranging from the requirement of sophisticated laboratories, need of skilled personnel, use of complex protocol, long wait times for results, and an overall high cost per test. These limitations have inspired researchers to search for alternative diagnostic methods that are fast, economical, and executable in low-resource laboratory settings. The discovery of Loop-mediated isothermal Amplification (LAMP) has provided a reliable substitute platform for the accurate detection of low copy number nucleic acids in the diagnosis of several viral diseases, including epidemics like Severe Acute Respiratory Syndrome (SARS) and Middle East Respiratory Syndrome (MERS). At present, a cocktail of LAMP assay reagents along with reverse transcriptase enzyme (Reverse Transcription LAMP, RT-LAMP) can be a robust solution for the rapid and cost-effective diagnosis for COVID-19, particularly in developing, and low-income countries. In summary, the development of RT-LAMP based diagnostic tools in a paper/strip format or the integration of this method into a microfluidic platform such as a Lab-on-a-chip may revolutionize the concept of PoCT for COVID-19 diagnosis. This review discusses the principle, technology and past research underpinning the success for using this method for diagnosing MERS and SARS, in addition to ongoing research, and the prominent prospect of RT-LAMP in the context of COVID-19 diagnosis.

Highlights

  • The emergence of a novel coronavirus disease (COVID-19) in late 2019 and its subsequent evolution into a pandemic created a severe global public health and economic concern

  • Severe Acute Respiratory Syndrome (SARS) coronavirus (SARS-CoV) is a zoonotic virus that reportedly appeared for the first time in Guangdong province, China, in 2003, which was transmitted to humans from bats, and eventually spread to 26 countries by infecting about 8000 people in the same year [54]

  • The first report using primers targeting nucleocapsid nucleotide sequences of Middle East Respiratory Syndrome (MERS) coronavirus implied that RT-Loop-mediated isothermal Amplification (LAMP) should be chosen over RT-Polymerase Chain Reaction (PCR) for rapid diagnosis due to better sensitivity

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Summary

Introduction

The emergence of a novel coronavirus disease (COVID-19) in late 2019 and its subsequent evolution into a pandemic created a severe global public health and economic concern. PCR-based methods require time-consuming and complicated protocols that limit their diagnostic efficacy in an active pandemic situation with rapidly and exponentially increasing number of incidences, in populous regions of the world Taken together, this necessitates substitution with an reliable molecular detection method for diagnosing and controlling COVID-19. On the diagnostic application of LAMP-based assays in past viral epidemics and the current COVID19 pandemic It provides notes on the basic principle, procedures, and possible improvements in the existing LAMP-based assays and stresses on its advantages over other nucleic acid amplification-based methods. At the end of the reaction, amplicons were mixed with the dye and the color changes indicated the presence of a positive reaction

LAMP for the Diagnosis of Viral Diseases
RT-LAMP for the Diagnosis of SARS
RT-LAMP for the Diagnosis of MERS
RT-LAMP for the Diagnosis of COVID-19
Challenges and Prospects
Schematic
Conclusions
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