National Centre for Foreign Animal Disease, Canadian Food Inspection Agency, Winnipeg, MB, Canada.
Sunnybrook Health Sciences Centre, Toronto, ON, Canada.
Sci Rep. 2021 Apr 30;11(1):9387. doi: 10.1038/s41598-021-88625-6.
The coronavirus disease 2019 (COVID-19) pandemic, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), calls for prompt and accurate diagnosis and rapid turnaround time for test results to limit transmission. Here, we evaluated two independent molecular assays, the Biomeme SARS-CoV-2 test, and the Precision Biomonitoring TripleLock SARS-CoV-2 test on a field-deployable point-of-care real-time PCR instrument, Franklin three9, in combination with Biomeme M1 Sample Prep Cartridge Kit for RNA 2.0 (M1) manual extraction system for rapid, specific, and sensitive detection of SARS-COV-2 in cell culture, human, and animal clinical samples. The Biomeme SARS-CoV-2 assay, which simultaneously detects two viral targets, the orf1ab and S genes, and the Precision Biomonitoring TripleLock SARS-CoV-2 assay that targets the 5' untranslated region (5' UTR) and the envelope (E) gene of SARS-CoV-2 were highly sensitive and detected as low as 15 SARS-CoV-2 genome copies per reaction. In addition, the two assays were specific and showed no cross-reactivity with Middle Eastern respiratory syndrome coronavirus (MERS-CoV), infectious bronchitis virus (IBV), porcine epidemic diarrhea virus (PEDV), transmissible gastroenteritis (TGE) virus, and other common human respiratory viruses and bacterial pathogens. Also, both assays were highly reproducible across different operators and instruments. When used to test animal samples, both assays equally detected SARS-CoV-2 genetic materials in the swabs from SARS-CoV-2-infected hamsters. The M1 lysis buffer completely inactivated SARS-CoV-2 within 10 min at room temperature enabling safe handling of clinical samples. Collectively, these results show that the Biomeme and Precision Biomonitoring TripleLock SARS-CoV-2 mobile testing platforms could reliably and promptly detect SARS-CoV-2 in both human and animal clinical samples in approximately an hour and can be used in remote areas or health care settings not traditionally serviced by a microbiology laboratory.
2019 年冠状病毒病（COVID-19）大流行是由严重急性呼吸系统综合征冠状病毒 2（SARS-CoV-2）引起的，需要快速准确的诊断和快速的检测结果周转时间，以限制传播。在这里，我们评估了两种独立的分子检测方法，即 Biomeme SARS-CoV-2 检测和 Precision Biomonitoring TripleLock SARS-CoV-2 检测，它们都在现场部署的即时护理实时 PCR 仪器 Franklin three9 上进行，结合 Biomeme M1 Sample Prep Cartridge Kit for RNA 2.0（M1）手动提取系统，用于快速、特异性和敏感地检测细胞培养物、人类和动物临床样本中的 SARS-COV-2。Biomeme SARS-CoV-2 检测同时检测两个病毒靶标，即orf1ab 和 S 基因，以及 Precision Biomonitoring TripleLock SARS-CoV-2 检测，该检测针对 SARS-CoV-2 的 5'非翻译区（5'UTR）和包膜（E）基因，检测灵敏度高达每个反应 15 个 SARS-CoV-2 基因组拷贝。此外，这两种检测方法均具有特异性，与中东呼吸综合征冠状病毒（MERS-CoV）、传染性支气管炎病毒（IBV）、猪流行性腹泻病毒（PEDV）、传染性胃肠炎（TGE）病毒和其他常见的人类呼吸道病毒和细菌病原体均无交叉反应。此外，这两种检测方法在不同操作人员和仪器上的重现性都很高。当用于测试动物样本时，两种检测方法均能同等检测到感染 SARS-CoV-2 的仓鼠拭子中的 SARS-CoV-2 遗传物质。M1 裂解缓冲液可在室温下在 10 分钟内完全使 SARS-CoV-2 失活，从而能够安全处理临床样本。总之，这些结果表明，Biomeme 和 Precision Biomonitoring TripleLock SARS-CoV-2 移动检测平台可以在大约一个小时内可靠且快速地检测人类和动物临床样本中的 SARS-CoV-2，并且可以在传统上未由微生物学实验室服务的偏远地区或保健环境中使用。
