Development and Evaluation of a Rapid GII Norovirus Detection Method Based on CRISPR-Cas12a.

Polish journal of microbiology Pub Date : 2024-03-04 eCollection Date: 2024-03-01 DOI:10.33073/pjm-2024-009
Xinyi Hu, Pei He, Tong Jiang, Jilu Shen
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Abstract

Norovirus is highly infectious and rapidly transmissible and represents a major pathogen of sporadic cases and outbreaks of acute gastroenteritis worldwide, causing a substantial disease burden. Recent years have witnessed a dramatic increase in norovirus outbreaks in China, significantly higher than in previous years, among which GII norovirus is the predominant prevalent strain. Therefore, rapid norovirus diagnosis is critical for clinical treatment and transmission control. Hence, we developed a molecular assay based on RPA combined with the CRISPER-CAS12a technique targeting the conserved region of the GII norovirus genome, the results of which could be displayed by fluorescence curves and immunochromatographic lateral-flow test strips. The reaction only required approximately 50 min, and the results were visible by the naked eye with a sensitivity reaching 102 copies/μl. Also, our method does not cross-react with other common pathogens that cause intestinal diarrhea. Furthermore, this assay was easy to perform and inexpensive, which could be widely applied for detecting norovirus in settings including medical institutions at all levels, particularly township health centers in low-resource areas.

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基于 CRISPR-Cas12a 的 GII 诺如病毒快速检测方法的开发与评估。
诺如病毒具有高度传染性和快速传播性,是全球零星病例和急性肠胃炎暴发的主要病原体,造成了巨大的疾病负担。近年来,我国诺如病毒暴发疫情急剧增加,明显高于往年,其中以 GII 型诺如病毒为主。因此,快速诊断诺如病毒对临床治疗和传播控制至关重要。因此,我们开发了一种基于 RPA 结合 CRISPER-CAS12a 技术的分子检测方法,以 GII 诺如病毒基因组的保守区为目标,其结果可通过荧光曲线和免疫层析侧流试纸显示。反应仅需约 50 分钟,结果肉眼可见,灵敏度达到 102 拷贝/μl。此外,我们的方法不会与其他导致肠道腹泻的常见病原体发生交叉反应。此外,该检测方法操作简便、成本低廉,可广泛应用于各级医疗机构,尤其是资源匮乏地区的乡镇卫生院检测诺如病毒。
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