Wang Tao, Jiang Wenqian, Huang Zhiqing, Yuan Zhitao, Chen Zhiwei, Lin Jun
Institute of Applied Genomics, Fuzhou University, No.2 Xueyuan Road, Fuzhou, 350108, China.
College of Biological Science and Engineering, Fuzhou University, Fuzhou, China.
Virol J. 2025 Jun 12;22(1):192. doi: 10.1186/s12985-025-02765-z.
Acute respiratory infections, caused by RNA viruses like respiratory syncytial virus, influenza, rhinovirus, and coronavirus, are major global health threats. Real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR) is the gold standard for detecting these viruses but is time-consuming, complex, and requires specialized equipment. There is a need for rapid, convenient, and multi-target detection methods to improve disease prevention and control. This study developed a multi-target immunochromatographic detection method using LbuCas13a protein and "band elimination" test strips for detecting SARS-CoV-2 and influenza virus. The method's performance was evaluated by testing known 5 positive and 4 negative samples for SARS-CoV-2 and comparing results with fluorescent PCR and colloidal gold methods. Detection sensitivity was quantified using digital PCR and qPCR. The immunochromatographic test strips showed 100% concordance with fluorescent PCR and colloidal gold methods in initial clinical SARS-CoV-2 detection. Subsequently, we used dual-target immunochromatographic test strips to detect 9 SARS-CoV-2 positive samples and 9 H3N2 positive samples. However, false negatives were observed in dual-target detection of SARS-CoV-2 and H3N2 samples, likely due to low sample concentration or sample degradation. The method had a minimum detection limit of 381.75 copies/µL, as determined by digital PCR and qPCR. The developed multi-target immunochromatographic detection method offers a rapid, low-cost, and simple approach for detecting both SARS-CoV-2 and influenza viruses. With high sensitivity, specificity, and reliability, this method holds promise as a practical tool for RNA virus diagnosis and improving public health response to respiratory infections.
由呼吸道合胞病毒、流感病毒、鼻病毒和冠状病毒等RNA病毒引起的急性呼吸道感染是全球主要的健康威胁。实时定量逆转录聚合酶链反应（RT-qPCR）是检测这些病毒的金标准，但耗时、复杂且需要专业设备。因此，需要快速、便捷且能多靶点检测的方法来改善疾病的预防和控制。本研究开发了一种使用LbuCas13a蛋白和“条带消除”试纸条的多靶点免疫层析检测方法，用于检测新型冠状病毒和流感病毒。通过检测已知的5份新型冠状病毒阳性样本和4份阴性样本，并将结果与荧光PCR和胶体金方法进行比较，对该方法的性能进行了评估。使用数字PCR和qPCR对检测灵敏度进行了定量分析。在最初的新型冠状病毒临床检测中，免疫层析试纸条与荧光PCR和胶体金方法的检测结果一致性达100%。随后，我们使用双靶点免疫层析试纸条检测了9份新型冠状病毒阳性样本和9份H3N2阳性样本。然而，在新型冠状病毒和H3N2样本的双靶点检测中观察到假阴性结果，这可能是由于样本浓度低或样本降解所致。通过数字PCR和qPCR测定，该方法的最低检测限为381.75拷贝/微升。所开发的多靶点免疫层析检测方法为新型冠状病毒和流感病毒的检测提供了一种快速、低成本且简便的方法。该方法具有高灵敏度、特异性和可靠性，有望成为RNA病毒诊断以及改善公共卫生对呼吸道感染应对能力的实用工具。
