BioFrontiers Institute, University of Colorado Boulder, Boulder, United States.
Department of Molecular, Cellular and Developmental Biology, University of Colorado Boulder, Boulder, United States.
Elife. 2021 Mar 29;10:e65113. doi: 10.7554/eLife.65113.
Here, we develop a simple molecular test for SARS-CoV-2 in saliva based on reverse transcription loop-mediated isothermal amplification. The test has two steps: (1) heat saliva with a stabilization solution and (2) detect virus by incubating with a primer/enzyme mix. After incubation, saliva samples containing the SARS-CoV-2 genome turn bright yellow. Because this test is pH dependent, it can react falsely to some naturally acidic saliva samples. We report unique saliva stabilization protocols that rendered 295 healthy saliva samples compatible with the test, producing zero false positives. We also evaluated the test on 278 saliva samples from individuals who were infected with SARS-CoV-2 but had no symptoms at the time of saliva collection, and from 54 matched pairs of saliva and anterior nasal samples from infected individuals. The Saliva TwoStep test described herein identified infections with 94% sensitivity and >99% specificity in individuals with sub-clinical (asymptomatic or pre-symptomatic) infections.
在这里，我们开发了一种基于逆转录环介导等温扩增的简单唾液新冠病毒检测方法。该方法分为两步：（1）用稳定溶液加热唾液；（2）用引物/酶混合物孵育来检测病毒。孵育后，含有新冠病毒基因组的唾液样本会变成亮黄色。由于该检测法依赖于 pH 值，因此可能会对一些天然酸性的唾液样本产生假阳性。我们报告了独特的唾液稳定化方案，使 295 份健康唾液样本与该检测法兼容，没有产生任何假阳性。我们还在 278 份采集时无症状的新冠病毒感染者的唾液样本和 54 份来自感染个体的配对唾液和前鼻样本中评估了该检测法。本文所述的两步唾液检测法在无症状（或症状前）感染个体中，对感染的检测具有 94%的敏感性和 >99%的特异性。
