Portable and simultaneous detection of four respiratory pathogens through a microfluidic LAMP and real-time fluorescence assay†

IF 3.6 3区 化学 Q2 CHEMISTRY, ANALYTICAL Analyst Pub Date : 2024-09-04 DOI:10.1039/D4AN00748D
Junwen Liu, Zhi Zeng, Feina Li, Bo Jiang, You Nie, Guohao Zhang, Biao Pang, Lin Sun and Rongzhang Hao
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Abstract

Respiratory pathogen infections are seasonally prevalent and are likely to cause co-infections or serial infections during peak periods of infection. Since they often cause similar symptoms, simultaneous and on-site detection of respiratory pathogens is essential for accurate diagnosis and efficient treatment of these infectious diseases. However, molecular diagnostic techniques for multiple pathogens in this field are lacking. Herein, we developed a microfluidic LAMP and real-time fluorescence assay for rapid detection of multiple respiratory pathogens using a ten-channel microfluidic chip with pathogen primers pre-embedded in the chip reaction well. The microfluidic chip provided a closed reaction environment, effectively preventing aerosol contamination and improving the accuracy of the detection results. Its corresponding detection instrument could automatically collect and display the fluorescence curve in real time, which was more conducive to the interpretation of results. The results showed that the developed method could specifically recognize the nucleic acid of influenza A(H1N1), Mycoplasma pneumoniae, respiratory syncytial virus type A, and SARS-CoV-2 with low detection limits of 104 copies per mL or 103 copies per mL. The test results on clinical samples demonstrated that the developed method has high sensitivity (92.00%) and high specificity (100.00%) and even has the capability to differentiate mixed-infection samples. With simple operation and high detection efficiency, the present portable and simultaneous detection assay could significantly improve the efficiency of on-site detection of respiratory infectious diseases and promote the accurate treatment, efficient prevention and control of the diseases.

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通过微流控 LAMP 和实时荧光检测法同时检测四种呼吸道病原体。
呼吸道病原体感染是季节性流行病,在感染高峰期很可能引起合并感染或连续感染。由于它们通常会引起相似的症状,因此同时和现场检测呼吸道病原体对于准确诊断和有效治疗这些传染病至关重要。然而,该领域缺乏针对多种病原体的分子诊断技术。在此,我们开发了一种微流控 LAMP 和实时荧光检测方法,利用十通道微流控芯片快速检测多种呼吸道病原体,芯片反应孔中预埋了病原体引物。微流控芯片提供了封闭的反应环境,有效防止了气溶胶污染,提高了检测结果的准确性。其相应的检测仪器可自动采集并实时显示荧光曲线,更有利于结果的解读。结果表明,所开发的方法能特异性识别甲型 H1N1 流感病毒、肺炎支原体、甲型呼吸道合胞病毒和 SARS-CoV-2 的核酸,检出限低,分别为 104 拷贝/毫升和 103 拷贝/毫升。对临床样本的检测结果表明,所开发的方法具有高灵敏度(92.00%)和高特异性(100.00%),甚至还能区分混合感染样本。该方法操作简单、检测效率高,可显著提高呼吸道传染病的现场检测效率,促进疾病的准确治疗和有效防控。
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来源期刊
Analyst
Analyst 化学-分析化学
CiteScore
7.80
自引率
4.80%
发文量
636
审稿时长
1.9 months
期刊介绍: The home of premier fundamental discoveries, inventions and applications in the analytical and bioanalytical sciences
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