Evaluation of the analytical performance of the 15-minute point-of-care DASH™ SARS-CoV-2 RT-qPCR test

Matthew A. Butzler; Jennifer L. Reed; Kirsten M. Knapton; Tania Afzal; Abhishek K. Agarwal; Jakob Schaeffer; Neeraj Saraiya; Lisa Oti; Ezekiel R. White; Emilie Giacobbe; Lacy M. Simons; Egon Anderson Ozer; Sally M. McFall

doi:10.1016/j.diagmicrobio.2023.116120

Evaluation of the analytical performance of the 15-minute point-of-care DASH™ SARS-CoV-2 RT-qPCR test

Matthew A. Butzler, Jennifer L. Reed, Kirsten M. Knapton, Tania Afzal, Abhishek K. Agarwal, Jakob Schaeffer, Neeraj Saraiya, Lisa Oti, Ezekiel R. White, Emilie Giacobbe, Lacy M. Simons, Egon Anderson Ozer, Sally M. McFall^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

2 Scopus citations

Abstract

Accurate and timely diagnosis for COVID-19 diagnosis allows highly effective antiviral medications to be prescribed. The DASH™ Rapid PCR System is a sample-to-answer point-of-care platform combining state-of-the-art PCR kinetics with sequence specific hybridization. The platform's first assay, the DASH™ SARS-CoV-2/S test for anterior nares direct swab specimens, received FDA Emergency Use Authorization in March 2022 for point-of-care use. Here we report the analytical characteristics of the assay including limit of detection, dynamic range, and robustness of SARS-CoV-2 variant detection. The limit of detection was determined by testing swabs contrived with one hundred copies of wild type or Omicron BA.5 virus and detecting 20/20 and 19/20, respectively. The dynamic range was assessed with contrived swabs containing 10²-10⁶ copies; the log-linear relationship between Cq and copy input was plotted, and the qPCR efficiency calculated from the slope of the line was 101.4%. Detection of seven SARS-CoV-2 variants was demonstrated.

Original language	English (US)
Article number	116120
Journal	Diagnostic Microbiology and Infectious Disease
Volume	108
Issue number	1
DOIs	https://doi.org/10.1016/j.diagmicrobio.2023.116120
State	Published - Jan 2024

Funding

The authors would like to thank Westberg Innovation for mechanical engineering design and fabrication support for the DASH™ analyzer and test cartridge, the DiMonte Group for development of the electronics and firmware for the DASH™ analyzer and past lab members for their contributions to this study: T Conis, D Epiney, O Johnson, D Patel, R Avila, M Kirichenko, R Kim, R Ross-Shannon, R Lehmann, and I Oguike. Sample preparation remains a challenge for nucleic acid testing. The use of sequence-specific capture reduces the coextraction of qPCR inhibitors enabling the processing of complex specimens such as saliva, vaginal swabs, plasma, and whole blood. Minute Molecular was funded by the U.S. National Institutes of Health National Institute of Biomedical Imaging and Bioengineering Rapid Acceleration of Diagnostics program [24] to accelerate the commercialization of the DASH™ platform. To capitalize on this investment, an extensive assay menu addressing public health priorities is under development. This project has been funded in part with Federal funds from the National Institute of Biomedical Imaging and Engineering (NIBIB), National Institutes of Health, Department of Health and Human Services, under Contract No. 75N92021C00013, NIH U54 EB027049S NIBIB, Fogarty International Center and Office of AIDS Research, and a grant from Minute Molecular Diagnostics, a wholly owned subsidiary of Nuclein. The study sponsors played no role in the design of any of the studies, in the collection, analysis and interpretation of data; in the writing of the manuscript; nor in the decision to submit the manuscript for publication.

Keywords

COVID-19
Diagnosis
Point-of-Care
Quantitative PCR

ASJC Scopus subject areas

Microbiology (medical)
Infectious Diseases

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1016/j.diagmicrobio.2023.116120

Cite this

Butzler, M. A., Reed, J. L., Knapton, K. M., Afzal, T., Agarwal, A. K., Schaeffer, J., Saraiya, N., Oti, L., White, E. R., Giacobbe, E., Simons, L. M., Ozer, E. A., & McFall, S. M. (2024). Evaluation of the analytical performance of the 15-minute point-of-care DASH™ SARS-CoV-2 RT-qPCR test. Diagnostic Microbiology and Infectious Disease, 108(1), Article 116120. https://doi.org/10.1016/j.diagmicrobio.2023.116120

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abstract = "Accurate and timely diagnosis for COVID-19 diagnosis allows highly effective antiviral medications to be prescribed. The DASH{\texttrademark} Rapid PCR System is a sample-to-answer point-of-care platform combining state-of-the-art PCR kinetics with sequence specific hybridization. The platform's first assay, the DASH{\texttrademark} SARS-CoV-2/S test for anterior nares direct swab specimens, received FDA Emergency Use Authorization in March 2022 for point-of-care use. Here we report the analytical characteristics of the assay including limit of detection, dynamic range, and robustness of SARS-CoV-2 variant detection. The limit of detection was determined by testing swabs contrived with one hundred copies of wild type or Omicron BA.5 virus and detecting 20/20 and 19/20, respectively. The dynamic range was assessed with contrived swabs containing 102-106 copies; the log-linear relationship between Cq and copy input was plotted, and the qPCR efficiency calculated from the slope of the line was 101.4%. Detection of seven SARS-CoV-2 variants was demonstrated.",

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doi = "10.1016/j.diagmicrobio.2023.116120",

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Butzler, MA, Reed, JL, Knapton, KM, Afzal, T, Agarwal, AK, Schaeffer, J, Saraiya, N, Oti, L, White, ER, Giacobbe, E, Simons, LM, Ozer, EA & McFall, SM 2024, 'Evaluation of the analytical performance of the 15-minute point-of-care DASH™ SARS-CoV-2 RT-qPCR test', Diagnostic Microbiology and Infectious Disease, vol. 108, no. 1, 116120. https://doi.org/10.1016/j.diagmicrobio.2023.116120

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AU - Reed, Jennifer L.

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AU - Afzal, Tania

AU - Agarwal, Abhishek K.

AU - Schaeffer, Jakob

AU - Saraiya, Neeraj

AU - Oti, Lisa

AU - White, Ezekiel R.

AU - Giacobbe, Emilie

AU - Simons, Lacy M.

AU - Ozer, Egon Anderson

AU - McFall, Sally M.

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N2 - Accurate and timely diagnosis for COVID-19 diagnosis allows highly effective antiviral medications to be prescribed. The DASH™ Rapid PCR System is a sample-to-answer point-of-care platform combining state-of-the-art PCR kinetics with sequence specific hybridization. The platform's first assay, the DASH™ SARS-CoV-2/S test for anterior nares direct swab specimens, received FDA Emergency Use Authorization in March 2022 for point-of-care use. Here we report the analytical characteristics of the assay including limit of detection, dynamic range, and robustness of SARS-CoV-2 variant detection. The limit of detection was determined by testing swabs contrived with one hundred copies of wild type or Omicron BA.5 virus and detecting 20/20 and 19/20, respectively. The dynamic range was assessed with contrived swabs containing 102-106 copies; the log-linear relationship between Cq and copy input was plotted, and the qPCR efficiency calculated from the slope of the line was 101.4%. Detection of seven SARS-CoV-2 variants was demonstrated.

AB - Accurate and timely diagnosis for COVID-19 diagnosis allows highly effective antiviral medications to be prescribed. The DASH™ Rapid PCR System is a sample-to-answer point-of-care platform combining state-of-the-art PCR kinetics with sequence specific hybridization. The platform's first assay, the DASH™ SARS-CoV-2/S test for anterior nares direct swab specimens, received FDA Emergency Use Authorization in March 2022 for point-of-care use. Here we report the analytical characteristics of the assay including limit of detection, dynamic range, and robustness of SARS-CoV-2 variant detection. The limit of detection was determined by testing swabs contrived with one hundred copies of wild type or Omicron BA.5 virus and detecting 20/20 and 19/20, respectively. The dynamic range was assessed with contrived swabs containing 102-106 copies; the log-linear relationship between Cq and copy input was plotted, and the qPCR efficiency calculated from the slope of the line was 101.4%. Detection of seven SARS-CoV-2 variants was demonstrated.

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Evaluation of the analytical performance of the 15-minute point-of-care DASH™ SARS-CoV-2 RT-qPCR test

Abstract

Funding

Keywords

ASJC Scopus subject areas

UN SDGs

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