Evaluation of extraction-free RT-qPCR methods for SARS-CoV-2 diagnostics

doi:10.1007/s00705-021-05165-0

. 2021 Oct;166(10):2825-2828.

doi: 10.1007/s00705-021-05165-0. Epub 2021 Jul 24.

Evaluation of extraction-free RT-qPCR methods for SARS-CoV-2 diagnostics

Alexander Domnich¹, Vanessa De Pace¹, Beatrice M Pennati², Patrizia Caligiuri³, Serena Varesano¹, Bianca Bruzzone¹, Andrea Orsi^{1

2}

Affiliations

¹ Hygiene Unit, San Martino Policlinico Hospital, IRCCS for Oncology and Neurosciences, Largo Rosanna Benzi 10, 16132, Genoa, Italy.
² Department of Health Sciences (DISSAL), University of Genoa, Via Pastore 1, 16132, Genoa, Italy.
³ Hygiene Unit, San Martino Policlinico Hospital, IRCCS for Oncology and Neurosciences, Largo Rosanna Benzi 10, 16132, Genoa, Italy. patrizia.caligiuri@hsanmartino.it.

PMID: 34302551
PMCID: PMC8302966
DOI: 10.1007/s00705-021-05165-0

Evaluation of extraction-free RT-qPCR methods for SARS-CoV-2 diagnostics

Alexander Domnich et al. Arch Virol. 2021 Oct.

. 2021 Oct;166(10):2825-2828.

doi: 10.1007/s00705-021-05165-0. Epub 2021 Jul 24.

Authors

Alexander Domnich¹, Vanessa De Pace¹, Beatrice M Pennati², Patrizia Caligiuri³, Serena Varesano¹, Bianca Bruzzone¹, Andrea Orsi^{1

2}

Affiliations

¹ Hygiene Unit, San Martino Policlinico Hospital, IRCCS for Oncology and Neurosciences, Largo Rosanna Benzi 10, 16132, Genoa, Italy.
² Department of Health Sciences (DISSAL), University of Genoa, Via Pastore 1, 16132, Genoa, Italy.
³ Hygiene Unit, San Martino Policlinico Hospital, IRCCS for Oncology and Neurosciences, Largo Rosanna Benzi 10, 16132, Genoa, Italy. patrizia.caligiuri@hsanmartino.it.

PMID: 34302551
PMCID: PMC8302966
DOI: 10.1007/s00705-021-05165-0

Abstract

Extraction-based real-time reverse transcription quantitative polymerase chain reaction (RT-qPCR) is currently the "gold standard" in SARS-CoV-2 diagnostics. However, some extraction-free RT-qPCR techniques have recently been developed. In this study, we compared the sensitivity of traditional extraction-based, heated extraction-free, and unheated extraction-free RT-qPCR methods for SARS-CoV-2 detection in nasopharyngeal swabs from symptomatic individuals. The unheated extraction-free method showed perfect agreement with the standard extraction-based RT-qPCR. By contrast, the heat-treated technique was associated with an 8.2% false negativity rate. Unheated extraction-free RT-qPCR for the molecular diagnosis of SARS-CoV-2 is a valuable alternative to the traditional extraction-based methods and may accelerate turnaround times by about two hours.

PubMed Disclaimer

Conflict of interest statement

The authors declare no conflict of interest regarding this publication.

Figures

**Fig. 1**
Distribution of the cycle threshold values by method used and gene target

See this image and copyright information in PMC

Cited by

High Diagnostic Accuracy of a Novel Lateral Flow Assay for the Point-of-Care Detection of SARS-CoV-2.
Giberti I, Costa E, Domnich A, Ricucci V, De Pace V, Garzillo G, Guarona G, Icardi G. Giberti I, et al. Biomedicines. 2022 Jun 30;10(7):1558. doi: 10.3390/biomedicines10071558. Biomedicines. 2022. PMID: 35884863 Free PMC article.
Evaluation of STANDARD^TM M10 SARS-CoV-2 assay as a diagnostic tool for SARS-CoV-2 in nasopharyngeal or oropharyngeal swab samples.
Parakatselaki ME, Alexi G, Zafiropoulos A, Sourvinos G. Parakatselaki ME, et al. J Clin Virol Plus. 2023 Feb;3(1):100129. doi: 10.1016/j.jcvp.2022.100129. Epub 2022 Dec 13. J Clin Virol Plus. 2023. PMID: 36530947 Free PMC article.
Comparative Bioinformatic Analysis Reveals Conserved Regions in SARS-CoV-2 Genome for RAPID Pandemic Response.
Karpuj MV, Shaytov DR, Shemer-Avni Y, Gideon M, Frenkel ZM, Sivan S. Karpuj MV, et al. Int J Mol Sci. 2024 May 25;25(11):5764. doi: 10.3390/ijms25115764. Int J Mol Sci. 2024. PMID: 38891951 Free PMC article.
Effect of the 2020/21 season influenza vaccine on SARS-CoV-2 infection in a cohort of Italian healthcare workers.
Domnich A, Orsi A, Sticchi L, Panatto D, Dini G, Ferrari A, Ogliastro M, Boccotti S, De Pace V, Ricucci V, Bruzzone B, Durando P, Icardi G. Domnich A, et al. Vaccine. 2022 Mar 15;40(12):1755-1760. doi: 10.1016/j.vaccine.2022.02.013. Epub 2022 Feb 7. Vaccine. 2022. PMID: 35153098 Free PMC article.
Long follow-up of BNT162b2 mRNA vaccine in healthcare workers (2020-2022): A retrospective longitudinal SARS-CoV-2 serological surveillance.
De Pace V, Bruzzone B, Ricucci V, Calcavecchia N, Guarona G, Giberti I, Costa E, Ogliastro M, Galano B, Nigro N, Murgia D, Nanni L, Orsi A. De Pace V, et al. Hum Vaccin Immunother. 2023 Aug;19(2):2258632. doi: 10.1080/21645515.2023.2258632. Epub 2023 Sep 19. Hum Vaccin Immunother. 2023. PMID: 37724517 Free PMC article.

See all "Cited by" articles

References

1. Park M, Cook AR, Lim JT, Sun Y, Dickens BL. A systematic review of COVID-19 epidemiology based on current evidence. J Clin Med. 2020;9:967. doi: 10.3390/jcm9040967. - DOI - PMC - PubMed
1. Pradhan D, Biswasroy P, Kumar Naik P, Ghosh G, Rath G. A review of current interventions for COVID-19 prevention. Arch Med Res. 2020;51:363–374. doi: 10.1016/j.arcmed.2020.04.020. - DOI - PMC - PubMed
1. Tang YW, Schmitz JE, Persing DH, Stratton CW. Laboratory diagnosis of COVID-19: Current issues and challenges. J Clin Microbiol. 2020;58:e00512–e520. doi: 10.1128/jcm.00512-20. - DOI - PMC - PubMed
1. McArthur L, Sakthivel D, Ataide R, Chan F, Richards JS, Narh CA. Review of burden, clinical definitions, and management of COVID-19 cases. Am J Trop Med Hyg. 2020;103:625–638. doi: 10.4269/ajtmh.20-0564. - DOI - PMC - PubMed
1. Venter M, Richter K. Towards effective diagnostic assays for COVID-19: A review. J Clin Pathol. 2020;73:370–377. doi: 10.1136/jclinpath-2020-206685. - DOI - PubMed

Publication types

Actions

MeSH terms

Actions
Actions
Actions
Actions
Actions
Actions
Actions
Actions
Actions

Substances

Actions

LinkOut - more resources

Full Text Sources
Medical
- MedlinePlus Health Information
Miscellaneous
- NCI CPTAC Assay Portal

[1] Park M, Cook AR, Lim JT, Sun Y, Dickens BL. A systematic review of COVID-19 epidemiology based on current evidence. J Clin Med. 2020;9:967. doi: 10.3390/jcm9040967. - DOI - PMC - PubMed

[2] Park M, Cook AR, Lim JT, Sun Y, Dickens BL. A systematic review of COVID-19 epidemiology based on current evidence. J Clin Med. 2020;9:967. doi: 10.3390/jcm9040967. - DOI - PMC - PubMed

[3] Pradhan D, Biswasroy P, Kumar Naik P, Ghosh G, Rath G. A review of current interventions for COVID-19 prevention. Arch Med Res. 2020;51:363–374. doi: 10.1016/j.arcmed.2020.04.020. - DOI - PMC - PubMed

[4] Pradhan D, Biswasroy P, Kumar Naik P, Ghosh G, Rath G. A review of current interventions for COVID-19 prevention. Arch Med Res. 2020;51:363–374. doi: 10.1016/j.arcmed.2020.04.020. - DOI - PMC - PubMed

[5] Tang YW, Schmitz JE, Persing DH, Stratton CW. Laboratory diagnosis of COVID-19: Current issues and challenges. J Clin Microbiol. 2020;58:e00512–e520. doi: 10.1128/jcm.00512-20. - DOI - PMC - PubMed

[6] Tang YW, Schmitz JE, Persing DH, Stratton CW. Laboratory diagnosis of COVID-19: Current issues and challenges. J Clin Microbiol. 2020;58:e00512–e520. doi: 10.1128/jcm.00512-20. - DOI - PMC - PubMed

[7] McArthur L, Sakthivel D, Ataide R, Chan F, Richards JS, Narh CA. Review of burden, clinical definitions, and management of COVID-19 cases. Am J Trop Med Hyg. 2020;103:625–638. doi: 10.4269/ajtmh.20-0564. - DOI - PMC - PubMed

[8] McArthur L, Sakthivel D, Ataide R, Chan F, Richards JS, Narh CA. Review of burden, clinical definitions, and management of COVID-19 cases. Am J Trop Med Hyg. 2020;103:625–638. doi: 10.4269/ajtmh.20-0564. - DOI - PMC - PubMed

[9] Venter M, Richter K. Towards effective diagnostic assays for COVID-19: A review. J Clin Pathol. 2020;73:370–377. doi: 10.1136/jclinpath-2020-206685. - DOI - PubMed

[10] Venter M, Richter K. Towards effective diagnostic assays for COVID-19: A review. J Clin Pathol. 2020;73:370–377. doi: 10.1136/jclinpath-2020-206685. - DOI - PubMed

Save citation to file

Email citation

Add to Collections

Add to My Bibliography

Your saved search

Create a file for external citation management software

Your RSS Feed

Evaluation of extraction-free RT-qPCR methods for SARS-CoV-2 diagnostics

Affiliations

Evaluation of extraction-free RT-qPCR methods for SARS-CoV-2 diagnostics

Authors

Affiliations

Abstract

Conflict of interest statement

Figures

Similar articles

Cited by

References

Publication types

MeSH terms

Substances

LinkOut - more resources

Full Text Sources

Medical

Miscellaneous

Abstract

Conflict of interest statement

Figures

Similar articles

Cited by

References

Publication types

MeSH terms

Substances

Related information

LinkOut - more resources

Full Text Sources

Medical

Miscellaneous