背景:甲型和乙型流感病毒检测在流行病学监测和疾病管理中至关重要。快速准确的诊断技术对于及时的临床干预和爆发预防至关重要。量子点编码微球已广泛应用于免疫检测。量子点编码微球与流式细胞术的集成是一种完善的技术,可以快速分析。因此,建立基于流式细胞术量子点微球的甲型和乙型流感病毒抗原多重检测方法将有助于疾病诊断。
目的:建立基于流式细胞术量子点编码微球技术的甲型和乙型流感病毒抗原共检测方法,这构成了多种呼吸道病毒生物标志物检测的基础。
方法:使用不同的量子点编码微球偶联抗甲型和乙型流感的单克隆抗体。在流式细胞仪上分别和同时检测已知的甲型和乙型流感抗原,并对检测条件进行了优化,建立了甲型和乙型流感抗原共检测方法,用于临床样本中的检测。将结果与荧光定量聚合酶链反应(PCR)方法进行比较,以验证该方法的临床性能。
结果:该方法对甲型和乙型流感抗原的检测限分别为26.1和10.7pg/mL,分别,两者的范围为15.6至250000pg/mL。在临床样本评估中,该方法与荧光定量PCR方法有很好的相关性,积极的,负,总体达标率为57.4%,100%,71.6%,分别。
结论:建立了甲型和乙型流感病毒抗原定量检测的多重检测方法,其特点是高灵敏度,良好的特异性,和广泛的检测范围,是有希望的临床应用。
BACKGROUND: Influenza A and B virus detection is pivotal in epidemiological surveillance and disease management. Rapid and accurate diagnostic techniques are crucial for timely clinical intervention and outbreak prevention. Quantum dot-encoded microspheres have been widely used in immunodetection. The integration of quantum dot-encoded microspheres with flow cytometry is a well-established technique that enables rapid analysis. Thus, establishing a multiplex detection method for influenza A and B virus antigens based on flow cytometry quantum dot microspheres will help in disease diagnosis.
OBJECTIVE: To establish a codetection method of influenza A and B virus antigens based on flow cytometry quantum dot-encoded microsphere technology, which forms the foundation for the assays of multiple respiratory virus biomarkers.
METHODS: Different quantum dot-encoded microspheres were used to couple the monoclonal antibodies against influenza A and B. The known influenza A and B antigens were detected both separately and simultaneously on a flow cytometer, and the detection conditions were optimized to establish the influenza A and B antigen codetection method, which was utilized for their detection in clinical samples. The results were compared with the fluorescence quantitative polymerase chain reaction (PCR) method to validate the clinical performance of this method.
RESULTS: The limits of detection of this method were 26.1 and 10.7 pg/mL for influenza A and B antigens, respectively, which both ranged from 15.6 to 250000 pg/mL. In the clinical sample evaluation, the proposed method well correlated with the fluorescent quantitative PCR method, with positive, negative, and overall compliance rates of 57.4%, 100%, and 71.6%, respectively.
CONCLUSIONS: A multiplex assay for quantitative detection of influenza A and B virus antigens has been established, which is characterized by high sensitivity, good specificity, and a wide detection range and is promising for clinical applications.