Room-temperature-storable PCR mixes for SARS-CoV-2 detection
| Autor: | Shengxiang Ge, Jun Zhang, Zhongfu Chen, Jin Wang, Kunyu Yang, Ningshao Xia, Shiyin Zhang, Zhang Dongxu, Tingdong Li, Jiasu Xu, Xiaosong Su, Yingbin Wang, Zecheng Zhong |
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| Rok vydání: | 2020 |
| Předmět: |
030213 general clinical medicine
2019-20 coronavirus outbreak Coronavirus disease 2019 (COVID-19) Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) Clinical Biochemistry Pneumonia Viral Cold storage 030204 cardiovascular system & hematology medicine.disease_cause Polymerase Chain Reaction Article 03 medical and health sciences Betacoronavirus 0302 clinical medicine COVID-19 Testing medicine Humans Food science Dna viral Pandemics Coronavirus DNA Primers Chromatography Clinical Laboratory Techniques SARS-CoV-2 Temperature Residual moisture COVID-19 General Medicine Freeze Drying PCR Freeze-drying DNA Viral Environmental science Coronavirus Infections |
| Zdroj: | Clinical Biochemistry |
| ISSN: | 1873-2933 |
| Popis: | Highlights • Room-temperature-storable PCR mixes for SARS-CoV-2 detection were introduced. • The mixes were consistent with those of freshly prepared, wet reagents. • Neither fridge for storage nor device for cold chain transportation is required. Objectives A novel coronavirus (severe acute respiratory syndrome coronavirus 2, SARS-CoV-2) emerged in late 2019, causing an outbreak of pneumonia [coronavirus disease 2019 (COVID-19)] globally. Although the use of ready-made reaction mixes can enable more rapid PCR-based diagnosis of COVID-19, the need to transport and store these mixes at low temperatures presents challenges to already overburdened logistics networks. Methods Here, we present an optimized freeze-drying procedure that allows SARS-CoV-2 PCR mixes to be transported and stored at ambient temperatures, without loss of activity. Additive-supplemented PCR mixes were freeze-dried. The residual moisture of the freeze-dried PCR mixes was measured by Karl-Fischer titration. Results We found that the freeze-dried PCR mixes with ~1.2% residual moisture are optimal for storage, transport, and reconstitution. The sensitivity, specificity, and repeatability of the freeze-dried reagents were similar to those of freshly prepared, wet reagents. The freeze-dried mixes retained activity at room temperature (18 ~ 25 °C) for 28 days, and for 14 and 10 days when stored at 37 °C and 56 °C, respectively. Conclusion The uptake of this approach will ease logistical challenges faced by transport networks and make more cold storage space available at diagnosis and hospital laboratories. |
| Databáze: | OpenAIRE |
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