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Room-temperature-storable PCR mixes for SARS-CoV-2 detection
Jiasu Xu1, Jin Wang1, Zecheng Zhong1
1State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Public Health, Xiamen University, Xiamen 361102, China.
Clinical Biochemistry
|June 28, 2020
Summary
Freeze-drying SARS-CoV-2 PCR mixes enables ambient temperature storage and transport, overcoming logistical challenges for COVID-19 diagnosis. This method maintains reagent activity and diagnostic accuracy without requiring cold chains.
Area of Science:
- Molecular Biology
- Virology
- Diagnostic Technologies
Background:
- The global COVID-19 pandemic necessitates rapid and accessible diagnostic testing.
- Current PCR-based diagnostic methods for SARS-CoV-2 require cold chain logistics for reagent storage and transport, straining supply chains.
- Optimizing reagent stability is crucial for widespread and equitable deployment of diagnostic tests.
Purpose of the Study:
- To develop and optimize a freeze-drying procedure for SARS-CoV-2 PCR mixes.
- To assess the stability and performance of freeze-dried PCR reagents at ambient temperatures.
- To alleviate logistical burdens associated with cold storage and transportation of diagnostic reagents.
Main Methods:
- Freeze-drying of additive-supplemented PCR mixes.
- Karl Fischer titration to determine residual moisture content.
- Evaluation of freeze-dried reagent sensitivity, specificity, and repeatability compared to fresh reagents.
- Stability testing at various elevated temperatures (37°C and 56°C) and room temperature (18-25°C).
Main Results:
- Optimal freeze-drying conditions achieved with approximately 1.2% residual moisture.
- Freeze-dried reagents demonstrated comparable sensitivity, specificity, and repeatability to fresh reagents.
- Reagents maintained activity for 28 days at room temperature, 14 days at 37°C, and 10 days at 56°C.
Conclusions:
- Freeze-drying offers a viable solution for stabilizing SARS-CoV-2 PCR mixes for ambient temperature transport and storage.
- This approach significantly reduces reliance on cold chain infrastructure, easing logistical challenges.
- The optimized procedure enhances the accessibility and scalability of COVID-19 diagnostic testing globally.
