Lyophilization of premixed COVID‐19 diagnostic RT‐qPCR reactions enables stable long‐term storage at elevated temperature
Lyophilization of premixed COVID‐19 diagnostic RT‐qPCR reactions enables stable long‐term storage at elevated temperature
复制标题
预混的 COVID-19 诊断 RT-qPCR 反应的冻干可以在高温下稳定地长期储存
DOI：
10.1002/biot.202000572
复制
发表时间：
2021
影响因子：
4.7
通讯作者：
Jewett, Michael C.
中科院分区：
文献类型：
--
作者：
Hammerling, Michael J.;Warfel, Katherine F.;Jewett, Michael C.
针对SARS-CoV-2的逆转录酶-定量聚合酶链反应（RT-qPCR）诊断检测是全球检测基础设施的基石。然而，这些测试需要冷链运输来分发，以及熟练技术人员的劳动来组装反应并解释结果。降低运输和人工成本的策略可以帮助诊断测试规模扩大和应对COVID-19爆发，以及未来的爆发。在这项研究中，我们测试了实验室开发的和商业化的SARS-CoV-2诊断RT-qPCR混合物通过冻干在高温下稳定的能力。将完全组装的反应物冻干，并在环境温度或高温下储存长达一个月，随后测定其检测合成SARS-CoV-2 RNA稀释液的能力。在测试的混合物中，我们发现一种商业混合物在冻干后在环境温度下储存至少30天后仍能保持活性和灵敏度。我们还证明，二糖等冻干保护剂可以使冻干诊断反应在高温（高达50°C）下稳定至少30天。我们预计，将这些方法纳入SARS-CoV-2诊断测试将通过减少测试设施的劳动力和消除冷链运输的需要来改善测试流程。
Reverse transcriptase‐quantitative polymerase chain reaction (RT‐qPCR) diagnostic tests for SARS‐CoV‐2 are the cornerstone of the global testing infrastructure. However, these tests require cold‐chain shipping to distribute, and the labor of skilled technicians to assemble reactions and interpret the results. Strategies to reduce shipping and labor costs at the point‐of‐care could aid in diagnostic testing scale‐up and response to the COVID‐19 outbreak, as well as in future outbreaks.In this study we test both lab‐developed and commercial SARS‐CoV‐2 diagnostic RT‐qPCR mixes for the ability to be stabilized against elevated temperature by lyophilization. Fully assembled reactions were lyophilized and stored for up to a month at ambient or elevated temperature and were subsequently assayed for their ability to detect dilutions of synthetic SARS‐CoV‐2 RNA.Of the mixes tested, we show that one commercial mix can maintain activity and sensitivity after storage for at least 30 days at ambient temperature after lyophilization. We also demonstrate that lyoprotectants such as disaccharides can stabilize freeze‐dried diagnostic reactions against elevated temperatures (up to 50°C) for at least 30 days.We anticipate that the incorporation of these methods into SARS‐CoV‐2 diagnostic testing will improve testing pipelines by reducing labor at the testing facility and eliminating the need for cold‐chain shipping.
DOI：
10.1016/j.ejpb.2013.05.019
发表时间：
2013-10-01
影响因子：
4.9
作者：
Kasper, Julia Christina;Winter, Gerhard;Friess, Wolfgang
通讯作者：
Friess, Wolfgang
保护性共轭疫苗的按需生物制造。
影响因子：
13.6
作者：
Stark JC;Jaroentomeechai T;Moeller TD;Hershewe JM;Warfel KF;Moricz BS;Martini AM;Dubner RS;Hsu KJ;Stevenson TC;Jones BD;DeLisa MP;Jewett MC
通讯作者：
Jewett MC
影响因子：
3.7
作者：
Bhadra S;Pothukuchy A;Shroff R;Cole AW;Byrom M;Ellefson JW;Gollihar JD;Ellington AD
通讯作者：
Ellington AD