Multiple molecular diagnostic platforms of nucleic acid amplification and antigen testing were used to diagnose SARS-CoV-2 infection. The diagnostic tests used were reverse transcription PCR (in accordance with the protocol developed by the National Institute of Infectious Diseases in Japan17 (link)), the FilmArray Respiratory Panel 2.1 test with the FilmArray Torch system (bioMérieux, Marcy-l’Etoile, France)18 (link), the Xpert Xpress SARS-CoV-2 test using Cepheid GeneXpert (Cepheid, Sunnyvale, CA)19 (link), and the Lumipulse antigen test with the LUMIPULSE G600II system (Fujirebio, Inc., Tokyo, Japan)20 (link),21 (link). All tests were performed with material obtained from nasopharyngeal swabs immersed in viral transport medium (Copan, Murrieta, CA).
Lumipulse g600ii system
Manufactured by Fujirebio
Sourced in Japan
The LUMIPULSE G600II system is a fully automated immunoassay analyzer designed for clinical laboratory testing. The system utilizes chemiluminescent enzyme immunoassay (CLEIA) technology to perform quantitative and qualitative measurements of various analytes in biological samples.
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4 protocols using lumipulse g600ii system
1
Comprehensive SARS-CoV-2 Diagnostic Protocols
2
Automated Chemiluminescence Enzyme Immunoassay for CSF Biomarkers
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The new automated chemiluminescence enzyme immunoassay (CLEIA) method (Lumipulse G600II System, Fujirebio, Tokyo, Japan) was used for the measurement of Aβ42, Aβ40, T-tau and P-tau181 in the CSF samples, using respective Lumipulse assays (Lumipulse Aβ42, Lumipulse Aβ40, Lumipulse T-tau, Lumipulse P-tau181 Immunoreaction Cartridges with the same ELISA’s antibodies and the same Substrate, Diluent and Wash Solutions reagents, Fujirebio, Ghent, Belgium). All measures were performed in the same batch of samples in run and single of reagents, calibration and three different internal controls (High, Medium and Low Levels) were processed at the beginning to ensure that all measured values testing the correct immunoassay functionality and were in the right range.
For the calibration, a three-point master curve was used with a range respectively between 0 and 3409 pg/mL for Aβ42 and between 0 and 30,000 pg/mL for Aβ40, a range between 0 and 2250 pg/mL for T-tau and between 0 e 400 pg/mL for P-tau181. All reagents and samples were uploaded on the roundabout of the automatic platform and results of the assay were provided on touchscreen in real-time at the end of each sample’s assays, after transferable on printer and/or USB system. The LoD was 7.17 pg/mL for Aβ42, 2.78 pg/mL for Aβ40, 141 pg/mL for T-tau, 0.282 pg/mL for P-Tau181 according to the manufacturer.
For the calibration, a three-point master curve was used with a range respectively between 0 and 3409 pg/mL for Aβ42 and between 0 and 30,000 pg/mL for Aβ40, a range between 0 and 2250 pg/mL for T-tau and between 0 e 400 pg/mL for P-tau181. All reagents and samples were uploaded on the roundabout of the automatic platform and results of the assay were provided on touchscreen in real-time at the end of each sample’s assays, after transferable on printer and/or USB system. The LoD was 7.17 pg/mL for Aβ42, 2.78 pg/mL for Aβ40, 141 pg/mL for T-tau, 0.282 pg/mL for P-Tau181 according to the manufacturer.
3
Multimodal SARS-CoV-2 Detection Protocols
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Nasopharyngeal swab samples were collected using cotton swabs and placed in viral transport media (Copan Diagnostics, Murrieta, CA, USA). Multiple molecular diagnostic testing platforms, including SARS-CoV-2 quantitative reverse transcriptase-PCR in accordance with the protocol developed by the National Institute of Infectious Diseases in Japan (Shirato et al., 2020 (link)), FilmArray Respiratory Panel 2.1 with the FilmArray Torch system (bioMérieux, Marcy-l'Etoile, France) (Hirotsu et al., 2020a (link)), Xpert Xpress SARS-CoV-2 test using Cepheid GeneXpert (Cepheid, Sunnyvale, CA, USA) (Hirotsu et al., 2022 (link)), and Lumipulse antigen test with the LUMIPULSE G600II system (Fujirebio, Inc., Tokyo, Japan) were used to identify positive samples for this study (Hirotsu et al., 2021 (link); Hirotsu et al., 2020b ).
4
SARS-CoV-2 Antigen Quantification
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The SARS-CoV-2 antigen levels were determined quantitatively with the Lumipulse SARS-CoV-2 antigen test (Fujirebio, Inc., Tokyo, Japan) as previously described [12 (link)]. Briefly, 700 μL of the VTM were vortexed and centrifuged at 2000×g for 5 min. Aliquots (100 μL) of the supernatant were tested on the LUMIPULSE G600II system (Fujirebio). For samples with an antigen level > 5000 pg/mL, the samples were diluted with the kit diluent and tested again, and the antigen level was calculated based on the dilution factor. We judged the results according to the manufacturer’s instruction.
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