Fan Haiyang, Gong Yonglong, Chang Mengying, Gao Juan, Li Mengjia, Chen Siyu, Yang Ruoyi, Zhao Muxue, Cui Yali, Hui Wenli
The College of Life Science, Northwest University, Xi'an, China.
Xi'an Gaoxin Hospital, Xi'an, China.
Microb Biotechnol. 2025 Jun;18(6):e70169. doi: 10.1111/1751-7915.70169.
Seasonal and persistent outbreaks of influenza viruses represent a significant challenge to global public health. Rapid, convenient and accurate diagnosis methods of influenza viruses are crucial for timely treatment to mitigate morbidity and mortality during both seasonal epidemics and pandemics. However, current diagnostic tools often face limitations in speed, accuracy or complexity of result interpretation; there is a great need for more efficient detection technology for influenza virus, especially for use in resource-limited settings or during large-scale outbreaks. This study developed a dual 'RT-LAMP-LFA' detection technology with gold magnetic nanoparticles for influenza virus. This method can simultaneously detect influenza A and B genes as well as internal reference genes within 35 min, with a detection limit of 80 copies/mL. This is the first time the RNase P gene has been introduced into a gold magnetic nanoparticle lateral flow assay system as a quality control measure to monitor the entire sampling and amplification process in virus detection and reveals the effects of loop primer deficiencies on the stability of the dual 'RT-LAMP-LFA' detection technology. Using fluorescent PCR detection technology as a benchmark, the analysis of a total of 70 clinical samples demonstrated a 100% agreement rate, confirming the applicability and accuracy of the dual 'RT-LAMP-LFA' detection system. This dual 'RT-LAMP-LFA' detection technology offers a novel option for diagnostic technology in hierarchical medical testing, presenting significant social importance and broad application prospects.
流感病毒的季节性和持续性爆发对全球公共卫生构成了重大挑战。快速、便捷且准确的流感病毒诊断方法对于在季节性流行和大流行期间及时治疗以降低发病率和死亡率至关重要。然而,当前的诊断工具在速度、准确性或结果解读的复杂性方面常常面临限制;迫切需要更高效的流感病毒检测技术,尤其是在资源有限的环境中或大规模爆发期间使用的技术。本研究开发了一种用于流感病毒的带有金磁纳米颗粒的双重“RT-LAMP-LFA”检测技术。该方法能够在35分钟内同时检测甲型和乙型流感病毒基因以及内参基因,检测限为80拷贝/毫升。这是首次将核糖核酸酶P基因引入金磁纳米颗粒侧向流动分析系统作为质量控制措施,以监测病毒检测中的整个采样和扩增过程,并揭示环引物缺陷对双重“RT-LAMP-LFA”检测技术稳定性的影响。以荧光PCR检测技术为基准,对总共70份临床样本的分析显示符合率为100%,证实了双重“RT-LAMP-LFA”检测系统的适用性和准确性。这种双重“RT-LAMP-LFA”检测技术为分级医学检测中的诊断技术提供了一种新选择,具有重大的社会意义和广阔的应用前景。