表面等离子体共振传感器技术在检测甲型流感病毒中的应用研究  

Surface plasmon resonance biosensor for detecting influenza A virus in human

作　　者：滕娟[1] 徐云庆[2] 顾大勇[2] 史蕾[2] 刘春晓[2] 赵纯中[2] 杨燕秋[2] 孙秋香[1] 欧青叶[2] 季明辉[2] 

机构地区：[1]中山大学公共卫生学院,广东广州510080 [2]深圳国际旅行卫生保健中心

出　　处：《中国国境卫生检疫杂志》2011年第3期145-149,共5页Chinese Journal of Frontier Health and Quarantine

基　　金：国家自然科学基金项目(30972827);国家质检总局科研基金项目(2010IK212);深港创新圈计划(联合资助)项目(ZYB200907090128A);深圳市科技计划切块项目(HZ0907004)

摘　　要：目的建立一种基于表面等离子体共振传感器技术检测人类甲型流感病毒的方法,并进行初步诊断应用。方法分别设计基于甲型流感M基因的引物和探针,制备RT-PCR扩增产物,优化并确定基于传感技术的实验条件,建立高灵敏度和高特异度的表面等离子体共振传感器方法。结果缓冲液的盐离子强度和pH值是影响杂交效果的关键因素,对其优化后标准曲线参数良好,甲型流感病毒标准品的Ru值与其相应梯度稀释标准品浓度呈良好的线性关系(R2>0.99),同时,该传感器在200 mmol/L氯化钠的磷酸盐缓冲液中可以有效区分DNA的完全匹配与单碱基错配,以乙型流感探针作为质控,该探针并不与甲型流感扩增产物发生特异性杂交。结论成功建立了高灵敏度和高特异度的甲型流感病毒表面等离子体共振传感器检测方法,为临床诊断或感染病例提供了技术平台。Objective To establish a detection method for influenza-A virus based on surface plasmon resonance （SPR） biosensor. Methods Design respectively primers and probe of influenza A virus based on M gene and prepare RT-PCR product. A sensitive and specific SPR method was established and optimized. Results Salt ionic strength and pH of the buffer were key elements that influence hybridization. The standard curve showed that the reflectivity shifts and PCR product concentration had a good linear relationship （ R 2 0.99）. In addition, the prepared biosensor method can effectively discriminate perfect complementary and other three types of mismatch： base substitute, base insertion and base deletion in 200 mmol/L NaCl-PBS buffer. Conclusion The established SPR biosensor method is sensitive and specific, which could be used as a prospective assay for detecting influenza-A virus. This system offers a good platform for diagnosing other causative organism.

关 键 词：表面等离子共振传感技术 甲型流感病毒 检测 应用 

分 类 号：R511.7[医药卫生—内科学]