Qiu Yue, Lu Ling, Halven Amanda, Terrio Rachel, Yuldelson Sydney, Dougal Natalie, Galbo Filippo, Lu Andrew, Gao Dexiang, Blomquist Bob, Zevallos Jose P, Lu Shi-Long, Yao Xin, Harry Brian L
Department of Otolaryngology - Head & Neck Surgery, University of Colorado Anschutz Medical Campus, Aurora, CO 80045, USA.
Liquid Biopsy Laboratory, Summit Biolabs, Aurora, CO 80045, USA.
J Biotechnol Biomed. 2024;7(2):214-220. doi: 10.26502/jbb.2642-91280144. Epub 2024 May 23.
The COVID-19 pandemic introduced an urgent need for rapid and high-throughput testing for SARS-CoV-2. RNA extraction is a major bottleneck for RT-qPCR. We describe a semi-automated, extraction-free RT-qPCR assay for detection of SARS-CoV-2 in nasal swab and saliva samples on a single platform. With a limit of detection of 4 copies/mL, this laboratory developed test performed equivalently to established methods requiring nucleic acid extraction. Five technologists staffing two shifts per day (80 person-hours) processed more than 400,000 samples over 10 months. Patients opted to provide nasal swab samples (83.6%) more frequently than saliva (16.4%), creating the added challenge of producing swab collection kits. Real-world testing data indicated a higher frequency of SARS-CoV-2 detection in saliva (10.1%) compared to nasal swab (7.7%). This cost-effective and quickly scalable approach is suitable for pandemic preparedness planning related to surveillance and diagnostic testing.
新冠疫情引发了对快速且高通量的新冠病毒检测的迫切需求。RNA提取是逆转录定量聚合酶链反应（RT-qPCR）的主要瓶颈。我们描述了一种在单一平台上检测鼻拭子和唾液样本中新冠病毒的半自动、无需提取的RT-qPCR检测方法。该实验室开发的检测方法检测限为每毫升4拷贝，其性能与需要核酸提取的既定方法相当。五名技术人员每天两班倒（80人时），在10个月内处理了超过40万个样本。患者选择提供鼻拭子样本（83.6%）的频率高于唾液样本（16.4%），这给生产拭子采集试剂盒带来了额外挑战。实际检测数据表明，唾液中新冠病毒的检测频率（10.1%）高于鼻拭子（7.7%）。这种具有成本效益且可快速扩展的方法适用于与监测和诊断检测相关的大流行防范规划。
