Abstract:
The influenza virus is a pervasive pathogen that exhibits increased prevalence during colder seasons, resulting in a significant annual occurrence of infections. Notably, pharmaceutical interventions effective against influenza A strains often exhibit limited efficacy against influenza B variants. Against this backdrop, the need for innovative approaches to accurately and swiftly differentiate and detect influenza B becomes evident. Biosensors play a pivotal role in this detection process, offering rapid, specific, and sensitive identification of the virus, facilitating timely intervention and containment efforts. Oligonucleotide sequences targeting the conserved B/Victoria/2/87 influenza virus NP region were designed. Nasopharyngeal swabs were collected from patients suspected of influenza virus infection, and viral RNA was extracted. RNA quality was assessed through one-step PCR. cDNA synthesis was performed using random hexamers, and real-time PCR quantified the influenza genome. Gold nanoparticles were immobilized on a surface to immobilize the specific DNA probe, and electrochemical hybridization was electrochemically followed. The biosensor exhibited high selectivity and effective distinction of complementary sequences from mismatches and influenza virus cDNA genome. The biosensor successfully detected the influenza B virus genome in real samples. Non-influenza samples yielded no significant hybridization signals. The comparison between the results obtained from the biosensor and real-time PCR revealed full agreement of these methods. The biosensor utilized electrochemical detection of hybridization and proved effective in detecting the influenza B virus genome with high specificity, sensitivity, and selectivity. Comparative analysis with real-time PCR underscored the accuracy and potential applicability of the biosensor in rapid and specific virus detection. This innovative approach holds promise for future diagnostic and epidemiological applications in detecting influenza B virus and other pathogens.
摘要:
流感病毒是一种普遍的病原体,在寒冷的季节患病率增加,导致每年大量感染。值得注意的是,对甲型流感株有效的药物干预措施通常对乙型流感变种的疗效有限.在这种背景下,需要创新的方法来准确和迅速地区分和检测乙型流感。生物传感器在这一检测过程中起着举足轻重的作用,提供快速,具体,以及对病毒的敏感鉴定,促进及时干预和遏制努力。设计了靶向保守的B/Victoria/2/87流感病毒NP区的寡核苷酸序列。从怀疑有流感病毒感染的病人身上采集鼻咽拭子,并提取病毒RNA。通过一步PCR评估RNA质量。使用随机六聚体进行cDNA合成,和实时PCR定量流感基因组。将金纳米粒子固定在表面以固定特定的DNA探针,然后电化学杂交。生物传感器显示出高选择性和有效区分来自错配和流感病毒cDNA基因组的互补序列。该生物传感器成功检测到真实样本中的乙型流感病毒基因组。非流感样品没有产生显著的杂交信号。从生物传感器和实时PCR获得的结果之间的比较表明这些方法完全一致。生物传感器利用电化学检测杂交,并被证明在检测乙型流感病毒基因组具有高特异性,灵敏度,和选择性。与实时PCR的比较分析强调了生物传感器在快速和特异性病毒检测中的准确性和潜在适用性。这种创新方法为检测乙型流感病毒和其他病原体的未来诊断和流行病学应用提供了希望。