Weng Yuesong, Zhou Jiefeng, Shi Yubo
Department of Laboratory Medicine, Ningbo First Hospital, Ningbo, China.
Ningbo AJcore Biosciences Inc., Ningbo, China.
Ann Transl Med. 2022 Oct;10(19):1064. doi: 10.21037/atm-22-4295.
The global coronavirus disease 2019 (COVID-19) pandemic has caused immeasurable losses to society. An important part of disease prevention and control is to diagnose infected patients as soon as possible, and nucleic acid detection plays an important role in diagnosing diseases such as COVID-19. Reverse transcription-polymerase chain reaction (RT-PCR) is a common method for the diagnosis of COVID-19 infection, but the false negative rate of covid nucleic acid is currently very high.
In this paper, HEK293T cells and lentivirus were used to verify the effect of virus preservation solution, fluorescence method was used to verify the inactivation effect of virus preservation solution, real-time fluorescence PCR experiment was used to verify that the genetic material in virus particles was not destroyed, and protein gel electrophoresis was used to verify the integrity of virus particles in the preservation solution.
In this study, we developed a new virus preservation solution using homogeneous reaction technology that can immediately inactivate the virus within 5 minutes at room temperature and maintain the stability of viral nucleic acid for at least 7 days. Importantly, we found that while our solution differed from existing commercial preservatives that contain guanidinium, analyzing the cycle threshold (Ct) showed that it still maintained virion integrity.
These advantages could reduce false negatives and the risk of laboratory exposure to infection. In addition, our solution achieved room temperature preservation, which could aid in the transportation of samples. Therefore, this new sample preservation solution could provide significant benefits to the clinical detection of viruses such as COVID-19.
2019年全球冠状病毒病(COVID-19)大流行给社会造成了不可估量的损失。疾病预防控制的一个重要部分是尽快诊断出感染患者,而核酸检测在诊断COVID-19等疾病中发挥着重要作用。逆转录聚合酶链反应(RT-PCR)是诊断COVID-19感染的常用方法,但目前新冠核酸的假阴性率非常高。
本文利用人胚肾293T细胞和慢病毒验证病毒保存液的效果,采用荧光法验证病毒保存液的灭活效果,通过实时荧光PCR实验验证病毒颗粒中的遗传物质未被破坏,并用蛋白质凝胶电泳验证保存液中病毒颗粒的完整性。
在本研究中,我们采用均相反应技术开发了一种新型病毒保存液,该保存液能在室温下5分钟内立即灭活病毒,并使病毒核酸的稳定性至少维持7天。重要的是,我们发现虽然我们的保存液与现有的含胍商业防腐剂不同,但分析循环阈值(Ct)表明它仍能保持病毒粒子的完整性。
这些优点可以减少假阴性以及实验室感染暴露的风险。此外,我们的保存液实现了室温保存,这有助于样本的运输。因此,这种新型样本保存液可为COVID-19等病毒的临床检测带来显著益处。