Sagoe Kate Obaayaa, Kyama Mutinda Cleophas, Maina Naomi, Kamita Moses, Njokah Muturi, Thiong'o Kelvin, Kanoi Bernard N, Wandera Ernest Apondi, Ndegwa Davies, Kinyua Dickson Mwenda, Gitaka Jesse
Department of Molecular Biology and Biotechnology, Pan African University Institute for Basic Sciences, Technology and Innovation (PAUSTI), Nairobi P.O. Box 62000-00200, Kenya.
Department of Medical Laboratory Science, College of Health Sciences, Jomo Kenyatta University of Agriculture & Technology, Nairobi P.O. Box 62000-00200, Kenya.
Diagnostics (Basel). 2023 May 7;13(9):1644. doi: 10.3390/diagnostics13091644.
Globally, the emergence of the coronavirus disease (COVID-19) has had a significant impact on life. The need for ongoing SARS-CoV-2 screening employing inexpensive and quick diagnostic approaches is undeniable, given the ongoing pandemic and variations in vaccine administration in resource-constrained regions. This study presents results as proof of concept to use hybridization chain reaction (HCR) and clustered regularly interspaced short palindromic repeats (CRISPR)/Cas12a complex for detecting SARS-CoV-2. HCR hairpin probes were designed using the NUPACK web-based program and further used to amplify the SARS-CoV-2 N gene in archived nasopharyngeal samples. The results were visualized using agarose gels and CRISPR Cas12a-based lateral flow strips. The assay was evaluated using the gold standard, real-time polymerase chain reaction (RT-PCR), as recommended by the World Health Organization (WHO). The results show the comparative efficiency of HCR to RT-PCR. This study shows that HCR and CRISPR are viable alternatives for diagnosing SARS-CoV-2 in samples.
在全球范围内，冠状病毒病（COVID-19）的出现对生活产生了重大影响。鉴于当前的大流行以及资源有限地区疫苗接种情况的差异，采用廉价且快速的诊断方法持续进行严重急性呼吸综合征冠状病毒2（SARS-CoV-2）筛查的必要性是不可否认的。本研究展示了使用杂交链式反应（HCR）和规律成簇间隔短回文重复序列（CRISPR）/Cas12a复合物检测SARS-CoV-2的概念验证结果。HCR发夹探针使用基于网络的NUPACK程序设计，并进一步用于扩增存档鼻咽样本中的SARS-CoV-2 N基因。结果通过琼脂糖凝胶和基于CRISPR Cas12a的侧向流动试纸条进行可视化。按照世界卫生组织（WHO）的建议，使用金标准实时聚合酶链反应（RT-PCR）对该检测方法进行评估。结果显示了HCR与RT-PCR相比的效率。本研究表明，HCR和CRISPR是诊断样本中SARS-CoV-2的可行替代方法。
