Central Virology Laboratory, Ministry of Health, Chaim Sheba Medical Center, Tel-Hashomer, Ramat-Gan, Israel.
Faculty of Medicine, Technion-Israel Institute of Technology, Haifa, Israel.
PLoS One. 2021 Mar 30;16(3):e0249149. doi: 10.1371/journal.pone.0249149. eCollection 2021.
Conducting numerous, rapid, and reliable PCR tests for SARS-CoV-2 is essential for our ability to monitor and control the current COVID-19 pandemic. Here, we tested the sensitivity and efficiency of SARS-CoV-2 detection in clinical samples collected directly into a mix of lysis buffer and RNA preservative, thus inactivating the virus immediately after sampling. We tested 79 COVID-19 patients and 20 healthy controls. We collected two samples (nasopharyngeal swabs) from each participant: one swab was inserted into a test tube with Viral Transport Medium (VTM), following the standard guideline used as the recommended method for sample collection; the other swab was inserted into a lysis buffer supplemented with nucleic acid stabilization mix (coined NSLB). We found that RT-qPCR tests of patients were significantly more sensitive with NSLB sampling, reaching detection threshold 2.1±0.6 (Mean±SE) PCR cycles earlier then VTM samples from the same patient. We show that this improvement is most likely since NSLB samples are not diluted in lysis buffer before RNA extraction. Re-extracting RNA from NSLB samples after 72 hours at room temperature did not affect the sensitivity of detection, demonstrating that NSLB allows for long periods of sample preservation without special cooling equipment. We also show that swirling the swab in NSLB and discarding it did not reduce sensitivity compared to retaining the swab in the tube, thus allowing improved automation of COVID-19 tests. Overall, we show that using NSLB instead of VTM can improve the sensitivity, safety, and rapidity of COVID-19 tests at a time most needed.
对 SARS-CoV-2 进行大量、快速和可靠的 PCR 检测对于我们监测和控制当前 COVID-19 大流行的能力至关重要。在这里,我们测试了直接采集到裂解缓冲液和 RNA 稳定剂混合物中的临床样本中 SARS-CoV-2 的检测灵敏度和效率,从而在采样后立即使病毒失活。我们测试了 79 名 COVID-19 患者和 20 名健康对照者。我们从每个参与者收集了两个样本(鼻咽拭子):一个拭子插入装有病毒运输培养基(VTM)的试管中,遵循作为样本采集推荐方法的标准指南;另一个拭子插入裂解缓冲液中补充有核酸稳定剂混合物(简称 NSLB)。我们发现,使用 NSLB 采样进行 RT-qPCR 测试的灵敏度明显更高,与来自同一患者的 VTM 样本相比,达到检测阈值的时间提前了 2.1±0.6(Mean±SE)个 PCR 循环。我们表明,这种改进很可能是因为在提取 RNA 之前,NSLB 样本不会在裂解缓冲液中稀释。在室温下将 NSLB 样本再提取 RNA 72 小时后,不会影响检测的灵敏度,这表明 NSLB 允许在没有特殊冷却设备的情况下长时间保存样本。我们还表明,与保留拭子在管中相比,在 NSLB 中旋转拭子并丢弃它不会降低灵敏度,从而允许 COVID-19 测试的自动化程度提高。总的来说,我们表明,在最需要的时候,使用 NSLB 代替 VTM 可以提高 COVID-19 测试的灵敏度、安全性和快速性。