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Related Experiment Video

Updated: Nov 9, 2025

Author Spotlight: Advancing Pathogen Diagnostics with Standardized LAMP

Author Spotlight: Advancing Pathogen Diagnostics with Standardized LAMP

Published on: September 8, 2023

Resilient SARS-CoV-2 diagnostics workflows including viral heat inactivation.

Maria Jose Lista^1,2,3, Pedro M Matos^1,2,3, Thomas J A Maguire^1,4,3

¹King's College London Diagnostics Team at Guy's Campus (London, UK).

Medrxiv : the Preprint Server for Health Sciences

|April 14, 2021

Summary

This study offers accessible SARS-CoV-2 detection workflows using commercial kits and heat inactivation of samples. Protocols are available to enable widespread testing, even without specialized facilities.

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Two-Step Reverse Transcription Droplet Digital PCR Protocols for SARS-CoV-2 Detection and Quantification

Two-Step Reverse Transcription Droplet Digital PCR Protocols for SARS-CoV-2 Detection and Quantification

Published on: March 31, 2021

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Last Updated: Nov 9, 2025

Author Spotlight: Advancing Pathogen Diagnostics with Standardized LAMP

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Author Spotlight: A Pseudotype Virus System for Assessing Omicron Subvariants and Neutralizing Antibodies in SARS-CoV-2 Research

Author Spotlight: A Pseudotype Virus System for Assessing Omicron Subvariants and Neutralizing Antibodies in SARS-CoV-2 Research

Published on: September 8, 2023

Two-Step Reverse Transcription Droplet Digital PCR Protocols for SARS-CoV-2 Detection and Quantification

Two-Step Reverse Transcription Droplet Digital PCR Protocols for SARS-CoV-2 Detection and Quantification

Published on: March 31, 2021

Area of Science:

Molecular Biology
Virology
Clinical Diagnostics

Background:

Global demand for SARS-CoV-2 detection reagents necessitates accessible testing solutions.
Many laboratories lack the capacity for kit-free or manual SARS-CoV-2 RNA extraction and detection.
Developing standardized, implementable workflows is crucial for expanding testing capabilities.

Approach:

Compared commercial RNA extraction kits (QIAamp, RNAdvance, Mag-Bind) and one-step RT-qPCR reagents (FastVirus, Luna, qPCRBIO).
Evaluated primer-probe sets for SARS-CoV-2 N and RdRP genes, assessing sensitivity with varying viral loads.
Validated heat inactivation of nasopharyngeal swabs (70-90°C) for SARS-CoV-2 inactivation using plaque assays and qPCR sensitivity.

Key Points:

All tested RNA extraction methods yielded comparable results.
FastVirus and Luna reagents demonstrated the highest sensitivity in RT-qPCR.
Detection of the viral N gene was more reliable than RdRP, especially for low viral titers.
Heat treatment effectively inactivated SARS-CoV-2 (original and B.1.1.7 variants) with minimal impact on qPCR sensitivity.
SARS-CoV-2 testing can be performed in settings lacking high-containment (CL-3) facilities.

Conclusions:

Multiple SARS-CoV-2 testing pipelines using commercially available reagents were established and validated.
Heat inactivation of clinical samples offers a safe and effective method for sample preparation.
The study provides freely available protocols and SOPs to facilitate widespread implementation of SARS-CoV-2 detection.