Abstract
The World Health Organizations declaration of the COVID-19 pandemic was a milestone for the scientific community. The high transmission rate and the huge number of deaths, along with the lack of knowledge about the virus and the evolution of the disease, stimulated a relentless search for diagnostic tests, treatments, and vaccines. The main challenges were the differential diagnosis of COVID-19 and the development of specific, rapid, and sensitive tests that could reach all people. RT-PCR remains the gold standard for diagnosing COVID-19. However, new methods, such as other molecular techniques and immunoassays emerged. Also, the need for accessible tests with quick results boosted the development of point of care tests (POCT) that are fast, and automated, with high precision and accuracy. This assay reduces the dependence on laboratory conditions and mass testing of the population, dispersing the pressure regarding screening and detection. This review summarizes the advances in the diagnostic field since the pandemic started, emphasizing various laboratory techniques for detecting COVID-19. We reviewed the main existing diagnostic methods, as well as POCT under development, starting with RT-PCR detection, but also exploring other nucleic acid techniques, such as digital PCR, loop-mediated isothermal amplification-based assay (RT-LAMP), clustered regularly interspaced short palindromic repeats (CRISPR), and next-generation sequencing (NGS), and immunoassay tests, and nanoparticle-based biosensors, developed as portable instruments for the rapid standard diagnosis of COVID-19.
【저자키워드】 severe acute respiratory syndrome coronavirus 2, Coronavirus disease 2019, Coronaviruses, Severe acute respiratory syndrome, digital PCR, enzyme-linked immunosorbent assay, Next-generation sequencing, field-effect transistor, nucleic acid amplification tests, lateral flow biosensor, Middle East respiratory syndrome, localized surface plasmon resonance, COVID-19coronavirus disease 2019, SARS-CoV-2severe acute respiratory syndrome coronavirus 2, MERSMiddle East Respiratory Syndrome, SARSSevere Acute Respiratory Syndrome, CRISPRclustered regularly interspaced short palindromic repeats, clustered regularly interspaced short palindromic repeats, ACE2Angiotensin-converting enzyme II, Angiotensin-converting enzyme II, ANVISABrazilian National Health Surveillance Agency, Brazilian National Health Surveillance Agency, AuNIsGold nanoislands, Gold nanoislands, cdPCRChip-based, Chip-based, CLIAChemiluminescent immunoassays, Chemiluminescent immunoassays, CoVCoronaviruses, ddPCRDroplet-based, Droplet-based, dPCRDigital PCR, dsDNADouble-stranded DNA, Double-stranded DNA, ELISAEnzyme-linked immunosorbent assay, endoRNaseRNA-guided endoribonuclease, RNA-guided endoribonuclease, FETField-effect transistor, ICLateral-flow immunochromatography, Lateral-flow immunochromatography, ICTimmunochromatographic tests, immunochromatographic tests, LFBlateral flow biosensor, LSPRLocalized surface plasmon resonance, mdPCRMicrofluidic digital PCR, Microfluidic digital PCR, mRT-LAMPMultiplex reverse transcription loop-mediated isothermal amplification, Multiplex reverse transcription loop-mediated isothermal amplification, NAATNucleic acid amplification tests, NGSNext-generation sequencing, NSP1 −NSP1616 nonstructural proteins, 16 nonstructural proteins, POCTPoint of care tests, Point of care tests, RT-LAMPReverse transcription loop-mediated isothermal amplification, Reverse transcription loop-mediated isothermal amplification, RT-PCRReal-Time Reverse Transcription Polymerase Chain Reaction, Real-Time Reverse Transcription Polymerase Chain Reaction, RT-RAAReverse transcription–recombinase aided amplification, Reverse transcription–recombinase aided amplification, RT-RPAReverse transcription-recombinase polymerase amplification, Reverse transcription-recombinase polymerase amplification, sgRNASingle guide RNA, Single guide RNA, SPIAsSolid-phase immunoassays, Solid-phase immunoassays, ssRNASingle-stranded RNA, Single-stranded RNA, 【초록키워드】 COVID-19, Evolution, pandemic, Vaccines, knowledge, COVID-19 pandemic, point of care, NGS, diagnostic, RT-PCR, virus, Laboratory, CRISPR, immunoassay, nucleic acid, RT-LAMP, Accuracy, diagnostic tests, POCT, automated, Scientific community, molecular, diagnostic methods, differential diagnosis, RT-PCR detection, deaths, isothermal, World Health Organization, Precision, gold standard, diagnosing, transmission rate, stimulated, lack, the disease, condition, reduce, diagnosis of COVID-19, palindromic repeat, 【제목키워드】 COVID-19, point of care, recent,