Clinical and Laboratory Diagnosis of SARS-CoV-2, the Virus Causing COVID-19

Silva, Severino Jefferson Ribeiro da; Silva, Caroline Targino Alves da; Guarines, Klarissa Miranda; Mendes, Renata Pessôa Germano; Pardee, Keith; Kohl, Alain; Pena, Lindomar

doi:10.1021/acsinfecdis.0c00274

Cited by 66 publications

(81 citation statements)

References 195 publications

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“…Next-generation sequencing (NGS) has recently been widely used in detecting the infectious strain mutation and biomolecular pathogenesis in the SARS-CoV-2 infection [ 43 , 44 ]. In this study, a transcriptome analysis was further conducted to explore the possible variations of the gene expression patterns in retinal organoids and monolayer cultures before and after the SARS-CoV-2 pseudovirus infection.…”

Section: Resultsmentioning

confidence: 99%

Expression of Endogenous Angiotensin-Converting Enzyme 2 in Human Induced Pluripotent Stem Cell-Derived Retinal Organoids

Lai

Chou

Chien

et al. 2021

IJMS

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Angiotensin-converting enzyme 2 (ACE2) was identified as the main host cell receptor for the entry of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and its subsequent infection. In some coronavirus disease 2019 (COVID-19) patients, it has been reported that the nervous tissues and the eyes were also affected. However, evidence supporting that the retina is a target tissue for SARS-CoV-2 infection is still lacking. This present study aimed to investigate whether ACE2 expression plays a role in human retinal neurons during SARS-CoV-2 infection. Human induced pluripotent stem cell (hiPSC)-derived retinal organoids and monolayer cultures derived from dissociated retinal organoids were generated. To validate the potential entry of SARS-CoV-2 infection in the retina, we showed that hiPSC-derived retinal organoids and monolayer cultures endogenously express ACE2 and transmembrane serine protease 2 (TMPRSS2) on the mRNA level. Immunofluorescence staining confirmed the protein expression of ACE2 and TMPRSS2 in retinal organoids and monolayer cultures. Furthermore, using the SARS-CoV-2 pseudovirus spike protein with GFP expression system, we found that retinal organoids and monolayer cultures can potentially be infected by the SARS-CoV-2 pseudovirus. Collectively, our findings highlighted the potential of iPSC-derived retinal organoids as the models for ACE2 receptor-based SARS-CoV-2 infection.

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Section: Resultsmentioning

confidence: 99%

Expression of Endogenous Angiotensin-Converting Enzyme 2 in Human Induced Pluripotent Stem Cell-Derived Retinal Organoids

Lai

Chou

Chien

et al. 2021

IJMS

View full text Add to dashboard Cite

show abstract

“…RT-PCR-based molecular methods represent the gold standard techniques used worldwide to make a confirmatory diagnosis of COVID-19 infection ( 46 ). Since the complete sequencing of the SARS-CoV-2 genome ( 26 ), researchers of different countries have begun to design molecular primers and probes specific to SARS-CoV-2 RNA sequences in order to perform differential diagnosis between COVID-19 infections and other pathologies with similar symptoms, such as seasonal flu or bacterial infections ( 47 , 48 ). Of note, the whole genome sequence of SARS-CoV-2 is 29,903 bp in length containing the following functional elements: A poly-A cap of 50 bases, the open reading frame 1/ab (ORF1/ab) containing the coding sequences for the RNA-dependent RNA polymerase (RdRP), the spike proteins, the envelope proteins, the membrane and nucleocapsid proteins, and a poly-A tail of 30 bp ( 49 ).…”

Section: Rt-pcr-based Molecular Testsmentioning

confidence: 99%

Current and innovative methods for the diagnosis of COVID‑19 infection (Review)

et al. 2021

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The Coronavirus Disease 2019 (COVID-19) pandemic has forced the scientific community to rapidly develop highly reliable diagnostic methods in order to effectively and accurately diagnose this pathology, thus limiting the spread of infection. Although the structural and molecular characteristics of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) were initially unknown, various diagnostic strategies useful for making a correct diagnosis of COVID-19 have been rapidly developed by private research laboratories and biomedical companies. At present, rapid antigen or antibody tests, immunoenzymatic serological tests and molecular tests based on RT-PCR are the most widely used and validated techniques worldwide. Apart from these conventional methods, other techniques, including isothermal nucleic acid amplification techniques, clusters of regularly inter-spaced short palindromic repeats/Cas (CRISPR/Cas)-based approaches or digital PCR methods are currently used in research contexts or are awaiting approval for diagnostic use by competent authorities. In order to provide guidance for the correct use of COVID-19 diagnostic tests, the present review describes the diagnostic strategies available which may be used for the diagnosis of COVID-19 infection in both clinical and research settings. In particular, the technical and instrumental characteristics of the diagnostic methods used are described herein. In addition, updated and detailed information about the type of sample, the modality and the timing of use of specific tests are also discussed.

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“…In one research, the efficiency of three novel real-time reverse transcription-PCR (RT-PCR) assays targeting RNA-dependent RNA polymerase (RdRp)/helicase (Hel), spike (S) and nucleocapsid (N) SARS-CoV-2 genes was developed and compared. RNA polymerase (RdRp)/(Hel) assay found to be effective with no cross reactivity with other viruses among samples [27]. Some of the RT-PCR kits manufactured at International level along with sample requirement has been shown in Table 5.…”

Section: International Statusmentioning

confidence: 99%

Development of RT-PCR Based Diagnosis of SARS-CoV-2

Patankar¹,

Zambare²

2022

Biotechnology to Combat COVID-19

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In the 2020, COVID-19 pandemic disease created an havoc situation world widely and mainly caused by Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2). It has been challenging task for researchers, scientists and medico-pharmaceutical organisations to find out rapid and reliable diagnosis methods. Among the all testing services, a Reverse Transcription Polymerase Chain Reaction (RT-PCR) is the more accurate, rapid and authenticated molecular technique used for most of the diagnosis of major diseases. It has been a global priority to fix the rapid diagnosis method to combat against the pandemic COVID-19. Thus, the present chapter mainly focussing on the progress of RT-PCR method development though various processes of data collection on isolation of whole genome sequence, its primer and method designing. In this scenario, India suddenly become the global leader for vaccine development and hence the challenges and RT-PCR kit development in India and rest of the world has been be discussed. World wide many Government and private agencies and industries have taken an initiative for diagnosis of SARS-CoV-2 hence this chapter also summarised the scope of RT-PCR to combat pandemic situation in future.

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Clinical and Laboratory Diagnosis of SARS-CoV-2, the Virus Causing COVID-19

Cited by 66 publications

References 195 publications

Expression of Endogenous Angiotensin-Converting Enzyme 2 in Human Induced Pluripotent Stem Cell-Derived Retinal Organoids

Expression of Endogenous Angiotensin-Converting Enzyme 2 in Human Induced Pluripotent Stem Cell-Derived Retinal Organoids

Current and innovative methods for the diagnosis of COVID‑19 infection (Review)

Development of RT-PCR Based Diagnosis of SARS-CoV-2

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