DNA-CHA/GO检测微平台的构筑及其在肺炎支原体分子诊断中的研究

近年来，肺炎支原体（MP）的感染发病率呈上升趋势，严重威胁人体健康。寻求敏感性强、特异性高的实验室诊断方法，对MP早期诊断、指导临床用药、疾病转归及预后起到至关重要的作用。基于纳米材料的核酸生物传感器用于病原微生物分子诊断是目前重要的研究方向之一。本项目在获取MP 16S rRNA基因特异性保守DNA序列的基础上，通过催化发卡自组装（CHA）荧光信号的淬灭与恢复及扩增和传导作用，利用氧化石墨烯（GO）的吸附及荧光淬灭特性，构筑DNA-CHA/GO无酶扩增生物传感检测微平台；然后验证可行性，优化测试条件，分析性能并与其他方法进行比较；最终采用临床标本分析方法的实用性，进行方法学评价。该检测微平台预期具有高敏感性、特异性、低成本、准确性及早期诊断等优势，可以弥补目前其他临床诊断方法的不足，并为研制与开发新型MP诊断方法提供理论基础及材料支撑，相信会对强化MP感染防治起到积极的促进作用。

In recent years, the incidence of Mycoplasma pneumoniae (MP) infection is on the rise, which is a serious threat to human health. The laboratory diagnosis method with high sensitivity and specificity plays an important role in early diagnosis, guiding clinical medication, influencing prognosis of MP. Nucleic acid biosensor based on nanomaterials is one of the most important research directions for the molecular diagnosis of pathogenic microbes. In this work, we will first obtain the specific conservative DNA sequence from the MP 16S rRNA gene, then establishing a method with efficiently fluorescence quenching, recovery , signal amplification and conduction of catalyzed hairpin assembly (CHA), using characteristics of adsorption and fluorescence quenching of graphene oxide (GO), to construct a DNA-CHA/GO biosensor microplatform for detection of nucleic acid through an enzyme-free signal amplification strategy. Then we will verify the feasibility, optimize the testing conditions, analyze the performance and compare with other methods. Finally, we will evaluate the method through the practicability of clinical specimen analysis. The detection microplatform is expected to have the advantages of high sensitivity, specificity, low cost, accuracy and early diagnosis. It can overcome the shortcomings of other clinical diagnostic methods and provide a theoretical and material basis for the development of a new MP molecular diagnosis method. It may play a positive role in promoting the prevention and treatment of MP infection.