Journal: Frontiers in microbiology

Article Title: Optimization and Clinical Validation of Colorimetric Reverse Transcription Loop-Mediated Isothermal Amplification, a Fast, Highly Sensitive and Specific COVID-19 Molecular Diagnostic Tool That Is Robust to Detect SARS-CoV-2 Variants of Concern.

doi: 10.3389/fmicb.2021.713713

Figure Lengend Snippet: FIGURE 9 | Schematic representation of SARS-CoV-2 genome indicating the amplicons for the COVID-19 molecular diagnostics by RT-LAMP. Structural representation of SARS-CoV-2 virion shows the main particle parts. LAMP primer regions are indicated as previously reported (Baek et al., 2020; Ben-Assa et al., 2020; Butler et al., 2020; Chow et al., 2020; Dudley et al., 2020; Song et al., 2021; Ganguli et al., 2020; Huang et al., 2020; Lamb et al., 2020; Lu et al., 2020; Mohon et al., 2020; Park et al., 2020; Rabe and Cepko, 2020; Thi et al., 2020; Yan et al., 2020; Yu et al., 2020; Zhang et al., 2020a,b; Anahtar et al., 2021; Bhadra et al., 2021; Bokelmann et al., 2021; González-González et al., 2021). ORF, open reading frame; RdRp, RNA-dependent RNA polymerase; NSP, nonstructural protein. Schematic representation created using Snap Gene Viewer software version 5.0.7; N1, N2, and N3_CDC correspond to the amplicons for SARS-CoV-2 detection by RT-PCR. Created with biorender.com.

Article Snippet: 2019-SARS-CoV-2 Detection Reagent Kit (Eiken Chemical, Tokyo, Japan) Turbidity or fluorescence (+ calcein) 63◦C/18–60 min 100%/100% 20–110 copies/Rx 130 PLA General Hospital, Beijing, China Yan et al., 2020 iLACO Respiratory (not detailed) NI ORF1ab NI Yes NEB #M1800 Color 65◦C/ ≥20 min 89.9%/NI 10 copies/μL (detection threshold of 60 copies/μL); equivalent to 35–37 Ct in RT-PCR 248 Shenyang province, China Yu et al., 2020 Respiratory swabs (not detailed) VTM N gene and ORF1a NI Yes NEB #M1800 + SYTO-9 Color and fluorescence 65◦C/30 min NI 4.8 copies/μL 6 Wuhan Institute of Virology, China Zhang et al., 2020a Synthetic SARS-CoV-2 RNA NA N gene; E gene and As1e gene (ORF1a) Human actin B gene Yes NEB #M1800 + SYTO-9 Color and fluorescence 65◦C/20 min 87.5% 2 copies/μL NA NA Zhang et al., 2020b NEB #M1800, WarmStart Colorimetric RT-LAMP 2 × Master Mix; NEB #E1700, WarmStart LAMP kit (DNA and RNA); RdRp, RNA-dependent RNA polymerase (harbored by ORF1ab SARS-COV-2 genome region); NI, noninformed; NA, not applied; UTM, Universal Transport medium; VTM, viral transport medium, commonly containing, Hank’s balanced salt solution at pH 7.4 containing BSA (1%), amphotericin (15 μg/mL), penicillin G (100 units/mL), and streptomycin (50 μg/mL); Prot K, proteinase K; BD UVTM, Becton–Dickinson Universal Viral Transport Media system; SPS, sample preservation solution; QuantiFluor, Promega system for dsDNA quantification using a DNA intercalating dye; Prot.

Techniques: Software, Reverse Transcription Polymerase Chain Reaction

Journal: International Journal of Molecular Sciences

Article Title: Development of Metal Complexes for Treatment of Coronaviruses

doi: 10.3390/ijms23126418

Figure Lengend Snippet: Auranofin inhibits SARS-CoV-2 proliferation in living organisms. Huh7 infection is associated with SARS-CoV-2 for 2 h at a multitude of infected (MOI) from one before ever being incubated with 4 M auranofin or 0.1 DMSO. Cell lysates and culturing filtrates were obtained 24 and 48 h after transmission, and viral RNA rates were measured using RT-PCR using primers and probe targeting the SARS-CoV-2 N1 and SARS-CoV-2 N2 regions. After measuring, standardizing, and correcting the human RNA extracted form tumor cells, the viral RNA values per ug of total cellular RNA were determined. The outcomes were compared for all specific primers, revealing a significant drop in viral RNA following 24 and 48 h. SARS-CoV-2 transmissibility titers were obtained using a cell test.48 h after invasion, in culturing filtrates The data are the mean SEM of two separate tests performed in duplicate, t -test ** p < 0.001. Reprinted/adapted with permission from Ref. .

Article Snippet: Most of the drugs studied had a good binding affinity to the SARS-CoV-2 major protease (M pro ), which was equivalent to Remdesivir, according to molecular docking calculations.

Techniques: Infection, Incubation, Transmission Assay, Reverse Transcription Polymerase Chain Reaction

Journal: International Journal of Molecular Sciences

Article Title: Development of Metal Complexes for Treatment of Coronaviruses

doi: 10.3390/ijms23126418

Figure Lengend Snippet: Auranofin-treated cells have a dose-dependent decrease in SARS-CoV-2 RNA: Auranofin (0.1–10 μM) was applied to SARSCOV-2 infected Huh7 cells in repeated dilutions. RT-PCR was used to quantify viral RNA in cell pellets and culture supernatants using primers and probes targeting the SARS-CoV-2 N1. The data were graphed using a non-linear regression model (GraphPad software). Auranofin reduced virus replication in infected cells with an EC50 of about 1.4 μM. The 50% cytotoxic concentration was around 5.7 μM. The data represent two separate tests that were carried out in duplicate, with permission from .

Article Snippet: Most of the drugs studied had a good binding affinity to the SARS-CoV-2 major protease (M pro ), which was equivalent to Remdesivir, according to molecular docking calculations.

Techniques: Infection, Reverse Transcription Polymerase Chain Reaction, Software, Concentration Assay

Journal: International Journal of Molecular Sciences

Article Title: Development of Metal Complexes for Treatment of Coronaviruses

doi: 10.3390/ijms23126418

Figure Lengend Snippet: Auranofin therapy significantly inhibits the incidence of SARS-CoV-2-induced immune cells in living organisms: qRT-PCR was used to determine mRNA levels of IL-6, IL-1, TNF, and NF-kB at 24 and 48 h after transmission. Following correcting the GAPDH gene, the fold variation in tumor cells compared to baseline controls was determined. The data shows the mean ± SEM of two separate tests that were performed in parallel. ** p < 0.001. Reprinted/adapted with permission from Ref. .

Article Snippet: Most of the drugs studied had a good binding affinity to the SARS-CoV-2 major protease (M pro ), which was equivalent to Remdesivir, according to molecular docking calculations.

Techniques: Quantitative RT-PCR, Transmission Assay

Journal: Cell Reports

Article Title: Isolation and characterization of cross-neutralizing coronavirus antibodies from COVID-19+ subjects

doi: 10.1016/j.celrep.2021.109353

Figure Lengend Snippet: Neutralizing mAbs as pre-exposure prophylaxis in k18-hACE2 mice CV1-1, CV1-30, and CV2-75 were assessed to see whether they could confer protection in a mouse model. (A) Experimental timeline. (B) Number of PFUs in the lungs 2 days following challenge. Error bars indicate SD. (C) Viral RNA in lung tissue 2 days after challenge was measured by qPCR and normalized to GAPDH expression. Error bars indicate SD. (D) Kaplan-Meyer survival curve of the viral load/titer in the lungs of remaining mice comparing the various treatment groups. Statistics were determined by one-way ANOVA with Dunnett’s multiple comparison test. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: The following Taqman Primer/Probe sets (Thermo Fisher Scientific) were used in this study: Gapdh (Mm99999915_g1).

Techniques: Expressing, Comparison

Journal: Cell Reports

Article Title: Isolation and characterization of cross-neutralizing coronavirus antibodies from COVID-19+ subjects

doi: 10.1016/j.celrep.2021.109353

Figure Lengend Snippet:

Article Snippet: The following Taqman Primer/Probe sets (Thermo Fisher Scientific) were used in this study: Gapdh (Mm99999915_g1).

Techniques: Virus, Infection, Recombinant, Affinity Column, Purification, Staining, Cloning, Cell Isolation, Transfection, Luciferase, Reverse Transcription, Gene Expression, Nested PCR, Sequencing, Expressing, Plasmid Preparation, Software, Microscopy