Performance of the RT-LAMP-based eazyplex® SARS-CoV-2 as a novel rapid diagnostic test

doi:10.1016/j.jcv.2021.104817

. 2021 May:138:104817.

doi: 10.1016/j.jcv.2021.104817. Epub 2021 Apr 1.

Performance of the RT-LAMP-based eazyplex® SARS-CoV-2 as a novel rapid diagnostic test

Renate Egerer¹, Birgit Edel¹, Bettina Löffler¹, Andreas Henke², Jürgen Rödel³

Affiliations

¹ Institute of Medical Microbiology, Jena University Hospital, Jena, Germany.
² Section of Experimental Virology, Institute of Medical Microbiology, Jena University Hospital, Jena, Germany.
³ Institute of Medical Microbiology, Jena University Hospital, Jena, Germany. Electronic address: juergen.roedel@med.uni-jena.de.

PMID: 33836452
PMCID: PMC8015392
DOI: 10.1016/j.jcv.2021.104817

Performance of the RT-LAMP-based eazyplex® SARS-CoV-2 as a novel rapid diagnostic test

Renate Egerer et al. J Clin Virol. 2021 May.

. 2021 May:138:104817.

doi: 10.1016/j.jcv.2021.104817. Epub 2021 Apr 1.

Authors

Renate Egerer¹, Birgit Edel¹, Bettina Löffler¹, Andreas Henke², Jürgen Rödel³

Affiliations

¹ Institute of Medical Microbiology, Jena University Hospital, Jena, Germany.
² Section of Experimental Virology, Institute of Medical Microbiology, Jena University Hospital, Jena, Germany.
³ Institute of Medical Microbiology, Jena University Hospital, Jena, Germany. Electronic address: juergen.roedel@med.uni-jena.de.

PMID: 33836452
PMCID: PMC8015392
DOI: 10.1016/j.jcv.2021.104817

Abstract

Background: Diagnostic assays for severe acute respiratory syndrome Coronavirus-2 (SARS-CoV-2) that are easy to perform and produce fast results are essential for timely decision making regarding the isolation of contagious individuals.

Objective: We evaluated the CE-approved eazyplex® SARS-CoV-2, a ready-to-use real time RT-LAMP assay for identification of the SARS-CoV-2 N and ORF8 genes from swabs in less than 30 min without RNA extraction.

Study design: Oropharyngeal and nasal swabs from 100 positive and 50 negative patients were inoculated into 0.9 % saline and tested by NeuMoDx™ RT-PCR. An aliquot was diluted fivefold in Copan sputum liquefying (SL) solution and directly analyzed by eazyplex® SARS-CoV-2. In addition, 130 patient swabs were prospectively tested with both methods in parallel. Analytical sensitivity of the assay was determined using virus stock dilutions.

Results: Positive percent agreement (PPA) between the eazyplex® SARS-CoV-2 and RT-PCR was 74 % for samples with Ct values < 35. When using a Ct cut-off ≤ 28 the PPA increased to 97.4 %. In the prospective part of the study overall PPA of the eazyplex® kit was 66.7 % but increased to 100 % when only Ct values ≤ 28 were considered. There were no false positive results. The median time to positivity was 12.5 min for the N gene and 16.75 min for ORF8. Analytical sensitivity was 3.75 TCID₅₀/mL. 10⁵ virus copies/mL were reproducibly detected.

Conclusion: The eazyplex® SARS-CoV-2 is a rapid assay that accurately identifies samples with high viral loads. It may be useful for near-patient testing outside of a molecular diagnostic laboratory.

Keywords: POC; RT-LAMP; Rapid diagnostic test; SARS-CoV-2; Without RNA extraction.

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Conflict of interest statement

The authors declare no conflicts of interest.

Figures

**Fig. 1**
NeuMoDx™ RT-PCR values for eazyplex® RT-LAMP-positive (n = 90) and negative test results (n = 34) in clinical samples.

See this image and copyright information in PMC

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References

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1. Mostafa H.H., Lamson D.M., Uhteg K., et al. Multicenter evaluation of the NeuMoDx™ SARS-CoV-2 test. J. Clin. Virol. 2020;130 doi: 10.1016/j.jcv.2020.104583. 104583. - DOI - PMC - PubMed
1. Candel F.J., Barreiro P., San Román J., et al. Recommendations for use of antigenic tests in the diagnosis of acute SARS-CoV-2 infection in the second pandemic wave: attitude in different clinical settings. Rev. Esp. Quimioter. 2020;33:466–484. doi: 10.37201/req/120.2020. - DOI - PMC - PubMed
1. Liotti F.M., Menchinelli G., Lalle E., et al. Performance of a novel diagnostic assay for rapid SARS-CoV-2 antigen detection in nasopharynx samples. Clin. Microbiol. Infect. 2020 doi: 10.1016/j.cmi.2020.09.030. - DOI - PMC - PubMed

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[1] Brooks Z.C., Das S. COVID-19 testing: impact of prevalence, sensitivity, and specificity on patient risk and cost. Am. J. Clin. Pathol. 2020;154:575–584. doi: 10.1093/ajcp/aqaa141. - DOI - PMC - PubMed

[2] Brooks Z.C., Das S. COVID-19 testing: impact of prevalence, sensitivity, and specificity on patient risk and cost. Am. J. Clin. Pathol. 2020;154:575–584. doi: 10.1093/ajcp/aqaa141. - DOI - PMC - PubMed

[3] Mostafa H.H., Hardick J., Morehead E., et al. Comparison of the analytical sensitivity of seven commonly used commercial SARS-CoV-2 automated molecular assays. J. Clin. Virol. 2020;130 doi: 10.1016/j.jcv.2020.104578. 104578. - DOI - PMC - PubMed

[4] Mostafa H.H., Hardick J., Morehead E., et al. Comparison of the analytical sensitivity of seven commonly used commercial SARS-CoV-2 automated molecular assays. J. Clin. Virol. 2020;130 doi: 10.1016/j.jcv.2020.104578. 104578. - DOI - PMC - PubMed

[5] Mostafa H.H., Lamson D.M., Uhteg K., et al. Multicenter evaluation of the NeuMoDx™ SARS-CoV-2 test. J. Clin. Virol. 2020;130 doi: 10.1016/j.jcv.2020.104583. 104583. - DOI - PMC - PubMed

[6] Mostafa H.H., Lamson D.M., Uhteg K., et al. Multicenter evaluation of the NeuMoDx™ SARS-CoV-2 test. J. Clin. Virol. 2020;130 doi: 10.1016/j.jcv.2020.104583. 104583. - DOI - PMC - PubMed

[7] Candel F.J., Barreiro P., San Román J., et al. Recommendations for use of antigenic tests in the diagnosis of acute SARS-CoV-2 infection in the second pandemic wave: attitude in different clinical settings. Rev. Esp. Quimioter. 2020;33:466–484. doi: 10.37201/req/120.2020. - DOI - PMC - PubMed

[8] Candel F.J., Barreiro P., San Román J., et al. Recommendations for use of antigenic tests in the diagnosis of acute SARS-CoV-2 infection in the second pandemic wave: attitude in different clinical settings. Rev. Esp. Quimioter. 2020;33:466–484. doi: 10.37201/req/120.2020. - DOI - PMC - PubMed

[9] Liotti F.M., Menchinelli G., Lalle E., et al. Performance of a novel diagnostic assay for rapid SARS-CoV-2 antigen detection in nasopharynx samples. Clin. Microbiol. Infect. 2020 doi: 10.1016/j.cmi.2020.09.030. - DOI - PMC - PubMed

[10] Liotti F.M., Menchinelli G., Lalle E., et al. Performance of a novel diagnostic assay for rapid SARS-CoV-2 antigen detection in nasopharynx samples. Clin. Microbiol. Infect. 2020 doi: 10.1016/j.cmi.2020.09.030. - DOI - PMC - PubMed

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Performance of the RT-LAMP-based eazyplex® SARS-CoV-2 as a novel rapid diagnostic test

Affiliations

Performance of the RT-LAMP-based eazyplex® SARS-CoV-2 as a novel rapid diagnostic test

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Abstract

Conflict of interest statement

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