Computer Vision-aided CRISPR Diagnostics for the Detection of COVID-19

Pattaraporn Nimsamer, Oraphan Mayuramart, Aubin Samacoits, Naphat Chantaravisoot, Chajchawan Nakhakes, Suchada Suphanpayak, Natta Padungwattanachoke, Nutcha Leelarthaphin, Hathaichanok Huayhongthong, T. Pisitkun, S. Payungporn, P. Hannanta-anan
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Abstract

Surveillance testing is a key strategy to control the spread of COVID-19. Unlike the gold standard testing method, quantitative reverse transcription polymerase chain reaction (RT-qPCR), CRISPR diagnostics have recently become a more appealing alternative as they are proven to be faster, simpler, and more affordable. However, the current CRISPR diagnostic readouts are typically non-quantitative, making them error-prone and lacking crucial information of viral load. To further improve the CRISPR diagnostic method, we have developed a custom computer vision algorithm that works in complement to common transilluminators to process fluorescence images of the diagnostic samples, quantify their fluorescence signals, and assign the test results. Our analysis showed that the quantified fluorescence intensity was directly correlated to the sample viral load, useful information for transmissibility and disease severity. Verified through laboratory and clinical samples, our algorithm accurately discriminated the samples with the viral RNA as low as 6.25 copies/uL, and correctly classified nasopharyngeal swab (NP swab) samples with 100% accuracy. Our work serves as a potential technique to improve the accuracy of CRISPR diagnostics of COVID-19 and promote rapid testing vital to the containment of the ongoing pandemic.
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计算机视觉辅助CRISPR诊断检测COVID-19
监测检测是控制COVID-19传播的关键策略。与金标准检测方法,定量逆转录聚合酶链反应(RT-qPCR)不同,CRISPR诊断最近成为一种更有吸引力的替代方法,因为它们被证明更快,更简单,更实惠。然而,目前的CRISPR诊断读数通常是非定量的,这使得它们容易出错,并且缺乏病毒载量的关键信息。为了进一步改进CRISPR诊断方法,我们开发了一种定制的计算机视觉算法,该算法与常见的泛照器互补,处理诊断样品的荧光图像,量化其荧光信号,并分配测试结果。我们的分析表明,定量荧光强度与样本病毒载量直接相关,这是传播性和疾病严重程度的有用信息。通过实验室和临床样本验证,我们的算法能够准确区分病毒RNA低至6.25拷贝/uL的样本,并以100%的准确率正确分类鼻咽拭子(NP拭子)样本。我们的工作可以作为一种潜在的技术来提高CRISPR诊断COVID-19的准确性,并促进对遏制当前大流行至关重要的快速检测。
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