tetano
Editor, Senior Moderator
J Clin Lab Anal
. 2022 Jun 12;e24513.
doi: 10.1002/jcla.24513. Online ahead of print.
Development and evaluation of time-resolved fluorescent immunochromatographic assay for quantitative detection of SARS-CoV-2 spike antigen
Buzhou Xu[SUP] 1 [/SUP], Hao Tang[SUP] 1 2 [/SUP], Yiming Weng[SUP] 1 [/SUP], Valerie Sloane Jones[SUP] 2 [/SUP], Shuhong Luo[SUP] 1 2 [/SUP], Chih Yun Cho[SUP] 2 [/SUP], Yongping Lin[SUP] 3 [/SUP], Jianmin Fang[SUP] 1 2 [/SUP], Xuedong Song[SUP] 1 [/SUP], Ruo-Pan Huang[SUP] 1 2 3 4 [/SUP]
Affiliations
Abstract
Background: The spread of COVID-19 worldwide caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has necessitated efficient, sensitive diagnostic methods to identify infected people. We report on the development of a rapid 15-minute time-resolved fluorescent (TRF) lateral flow immunochromatographic assay for the quantitative detection of the SARS-CoV-2 spike protein receptor-binding domain (S1-RBD).
Objectives: Our objective was to develop an efficient method of detecting SARS-CoV-2 within 15 min of sample collection.
Methods: We constructed and evaluated a portable, disposable lateral flow device, which detected the S1-RBD protein directly in nasopharyngeal swab samples. The device emits a fluorescent signal in the presence of S1-RBD, which can be captured by an automated TRF instrument.
Results: The TRF lateral flow assay signal was linear from 0 to 20 ng/ml and demonstrated high accuracy and reproducibility. When evaluated with clinical nasopharyngeal swabs, the assay was performed at >80% sensitivity, >84% specificity, and > 82% accuracy for detection of the S1-RBD antigen.
Conclusion: The new S1-RBD antigen test is a rapid (15 min), sensitive, and specific assay that requires minimal sample preparation. Critically, the assay correlated closely with PCR-based methodology in nasopharyngeal swab samples, showing that the detected S1-RBD antigen levels correlate with SARS-CoV-2 virus load. Therefore, the new TRF lateral flow test for S1-RBD has potential application in point-of-care settings.
Keywords: COVID-19; RT-PCR; SARS-CoV-2; Spike protein; TRF lateral flow; time-resolved fluorescence.
. 2022 Jun 12;e24513.
doi: 10.1002/jcla.24513. Online ahead of print.
Development and evaluation of time-resolved fluorescent immunochromatographic assay for quantitative detection of SARS-CoV-2 spike antigen
Buzhou Xu[SUP] 1 [/SUP], Hao Tang[SUP] 1 2 [/SUP], Yiming Weng[SUP] 1 [/SUP], Valerie Sloane Jones[SUP] 2 [/SUP], Shuhong Luo[SUP] 1 2 [/SUP], Chih Yun Cho[SUP] 2 [/SUP], Yongping Lin[SUP] 3 [/SUP], Jianmin Fang[SUP] 1 2 [/SUP], Xuedong Song[SUP] 1 [/SUP], Ruo-Pan Huang[SUP] 1 2 3 4 [/SUP]
Affiliations
- PMID: 35692032
- DOI: 10.1002/jcla.24513
Abstract
Background: The spread of COVID-19 worldwide caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has necessitated efficient, sensitive diagnostic methods to identify infected people. We report on the development of a rapid 15-minute time-resolved fluorescent (TRF) lateral flow immunochromatographic assay for the quantitative detection of the SARS-CoV-2 spike protein receptor-binding domain (S1-RBD).
Objectives: Our objective was to develop an efficient method of detecting SARS-CoV-2 within 15 min of sample collection.
Methods: We constructed and evaluated a portable, disposable lateral flow device, which detected the S1-RBD protein directly in nasopharyngeal swab samples. The device emits a fluorescent signal in the presence of S1-RBD, which can be captured by an automated TRF instrument.
Results: The TRF lateral flow assay signal was linear from 0 to 20 ng/ml and demonstrated high accuracy and reproducibility. When evaluated with clinical nasopharyngeal swabs, the assay was performed at >80% sensitivity, >84% specificity, and > 82% accuracy for detection of the S1-RBD antigen.
Conclusion: The new S1-RBD antigen test is a rapid (15 min), sensitive, and specific assay that requires minimal sample preparation. Critically, the assay correlated closely with PCR-based methodology in nasopharyngeal swab samples, showing that the detected S1-RBD antigen levels correlate with SARS-CoV-2 virus load. Therefore, the new TRF lateral flow test for S1-RBD has potential application in point-of-care settings.
Keywords: COVID-19; RT-PCR; SARS-CoV-2; Spike protein; TRF lateral flow; time-resolved fluorescence.