A Portable RT-LAMP/CRISPR Machine for Rapid COVID-19 Screening.

Meysam Rezaei,Olga Shimoni,William Rawlinson,Majid Ebrahimi Warkiani,Dayong Jin,Sajad Razavi Bazaz,Dorsa Morshedi Rad

doi:10.3390/bios11100369

Abstract

The COVID-19 pandemic has changed people’s lives and has brought society to a sudden standstill, with lockdowns and social distancing as the preferred preventative measures. To lift these measurements and reduce society’s burden, developing an easy-to-use, rapid, and portable system to detect SARS-CoV-2 is mandatory. To this end, we developed a portable and semi-automated device for SARS-CoV-2 detection based on reverse transcription loop-mediated isothermal amplification followed by a CRISPR/Cas12a reaction. The device contains a heater element mounted on a printed circuit board, a cooler fan, a proportional integral derivative controller to control the temperature, and designated areas for 0.2 mL Eppendorf® PCR tubes. Our system has a limit of detection of 35 copies of the virus per microliter, which is significant and has the capability of being used in crisis centers, mobile laboratories, remote locations, or airports to diagnose individuals infected with SARS-CoV-2. We believe the current methodology that we have implemented in this article is beneficial for the early screening of infectious diseases, in which fast screening with high accuracy is necessary.

Highlights

The SARS-CoV-2 virus causes COVID-19, a serious and life-threatening disease with a wide spectrum of symptoms, ranging from fever and cough to dyspnea, and in severe cases, resulting in intensive care unit (ICU) admission of patients, organ failure, and death [1,2]
For the Biotin-tagged pnouscilteiivceascaidmsp(lMesi,letnwioa HlinyebsriwDeerteecot b1s,eTrvweidstDonx,tCheamlabterirdagl efl,oUwKs).trFiposr: thUencut reporters positive samplceasp, ttuwroedlinatesthwe efirrestolbinseer(vceodntornolt)haenldatceurtarlefploowrtesrtsricpasp: tUunrecdutartetpheorsteecrosncdapli-ne; while tured at the firsint lnineega(ctiovnetrsoalm) apnleds,count lryepoonretelirnseca(cpotnutrreodl)aotfthcaepsteucroenddulninceu(ttreespt)o; rwtehrislewians detected. negative samples, oAnflyteor nvealliidnaet(icnogntthroelf)eoafscibaiplittuyreodf ouunrcuapt prerpooacrtherfsorwdaestedcettiencgtetdh.e SARS-CoV-2 virus, we determined the limit of detection (LoD) in a controlled situation using lab-based instruments before the final design and production of our portable RT-LAMP/CRISPR
The copy number of each sample was confirmed and the LoD of commercial RT-qPCR assay for SARS-CoV-2 detection was determined using the cycle threshold (Ct) values (Figure 1C)

Summary

Introduction

The SARS-CoV-2 virus causes COVID-19, a serious and life-threatening disease with a wide spectrum of symptoms, ranging from fever and cough to dyspnea, and in severe cases, resulting in intensive care unit (ICU) admission of patients, organ failure, and death [1,2]. Negative samples, oAnflyteor nvealliidnaet(icnogntthroelf)eoafscibaiplittuyreodf ouunrcuapt prerpooacrtherfsorwdaestedcettiencgtetdh.e SARS-CoV-2 virus, we determined the limit of detection (LoD) in a controlled situation using lab-based instruments before the final design and production of our portable RT-LAMP/CRISPR

Results

Conclusion