Department of Medical Microbiology and Infection Control, Franciscus Gasthuis & Vlietland, Rotterdam, the Netherlands; Department of Epidemiology, Julius Center for Health Sciences and Primary Care, University Medical Center Utrecht, Utrecht, the Netherlands.
Department of Medical Microbiology and Infection Control, Franciscus Gasthuis & Vlietland, Rotterdam, the Netherlands.
J Clin Virol. 2021 Aug;141:104903. doi: 10.1016/j.jcv.2021.104903. Epub 2021 Jun 19.
Comprehensive and up-to-date monitoring of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants of concern (VOC) is crucial as these are characterized by their increased transmissibility, immune evasion and virulence.
To describe the wide-scale implementation of a reverse transcriptase polymerase chain reaction (RT-PCR) multiple variants assay with melting curve analysis as a routine procedure.
We prospectively performed multiple variants RT-PCR on consecutive SARS-CoV-2 RT-PCR positive samples from patients, healthcare workers and nursing home residents from our hospital catchment area. This technique was implemented in our automated Roche FLOW system with a turn-around time of 6 h.
Between February 1 and May 2, 2021, 989 samples were tested by the variant RT-PCR. Our method was validated by comparison of variant RT-PCR to whole genome sequencing testing. We observed an increase over time in the proportion of UK variant that became the dominant variant, and the concurrent emergence of the South-African and Brazilian variants. Prompt public health responses for infection control were possible because of this rapid screening method, resulting in early detection and reduction of unnoticed spread of VOC as early as possible.
A variant RT-PCR with additional melting curve analyses is a feasible, rapid and efficient screening strategy that can be implemented in routine microbiological laboratories.
全面且最新地监测严重急性呼吸综合征冠状病毒 2（SARS-CoV-2）变体是至关重要的，因为这些变体的特点是传染性增加、免疫逃逸和毒力增强。
描述一种广泛应用的逆转录聚合酶链反应（RT-PCR）多变体分析与熔解曲线分析作为常规程序。
我们前瞻性地对来自我们医院服务区的患者、医护人员和养老院居民的连续 SARS-CoV-2 RT-PCR 阳性样本进行了多变体 RT-PCR。该技术在我们的罗氏 FLOW 自动化系统中实施，周转时间为 6 小时。
2021 年 2 月 1 日至 5 月 2 日，共有 989 个样本通过变体 RT-PCR 进行了检测。我们的方法通过与全基因组测序测试的比较得到了验证。我们观察到随着时间的推移，英国变体的比例增加，成为主要变体，同时南非和巴西变体也出现了。由于这种快速筛选方法，可以及时进行公共卫生响应以进行感染控制，从而尽早发现和减少 VOC 的无察觉传播。
带有附加熔解曲线分析的变体 RT-PCR 是一种可行、快速且高效的筛选策略，可以在常规微生物学实验室中实施。
