SERS指纹谱用于志贺菌鉴定及喹诺酮药敏的快速检测新技术研究

Our two previous NSFC project (81000754, 81471994) research found that: Shigella infection and quinolone resistance is serious, however, the bacterium identification and drug sensitivity test need more than 3 days, which cannot meet the clinical needs. Surface-enhanced Raman spectroscopy (SERS) has the advantages of rapid, high sensitivity and fingerprint characteristic spectrum, but the analysis of clinical samples is at an exploratory stage, and it is not yet available for drug susceptibility testing. We innovatively applied SERS technology to the rapid detection of Shigella, and preliminary experiments confirmed that this novel technology can effectively distinguish different serotypes of Shigella. This project intends to further establish and optimize this new technology, and systematically evaluate its application value for the rapid identification of Shigella in clinical samples and the rapid detection of quinolone susceptibility.Firstly, prepare high-performance materials which are suitable for the separation of pathogenic bacteria and fingerprint signal enhancement in fecal samples, establish a standardized bacterial SERS fingerprint spectrum, and construct Shigella fingerprint database; Secondly, the PEI-modified magnetic nanoparticles were then used to isolate bacteria from the fecal samples and cultured through blood plate and quinolone resistant plates; Last, use SERS fingerprint from single colonies for accurate identification of Shigella different serotypes and resistant strains. This project will provide a novel and rapid technique for diagnosis and treatment of clinical Shigella infection.

申请人前两项国家自然科学基金(81000754、81471994)研究发现：志贺菌感染及对喹诺酮耐药形势严峻，但其检测与药敏需3天以上，无法满足临床需求。表面增强拉曼光谱(SERS)技术具有快速、高灵敏度和指纹特征光谱等优势，但对临床样本分析处于探索阶段，且目前尚不能用于药敏分析。我们创新性地将该技术用于志贺菌快速检测，预实验证实：该新技术可有效区分志贺菌不同血清型。本课题拟进一步建立与优化该新技术，并系统评估其用于临床样本中志贺菌快速鉴定及对喹诺酮药敏快速检测的应用价值。首先制备适合粪便标本中志贺菌分离及指纹信号增强的高性能材料，建立标准化SERS指纹谱，构建志贺菌指纹数据库；然后利用PEI修饰的磁性纳米颗粒从粪便中分离细菌并通过血平板及喹诺酮抗性平板培养；最后利用生长的单菌落的SERS指纹谱准确鉴定志贺菌不同血清型及耐药株。本课题的完成，将为临床志贺菌感染诊疗提供一种快速检测的新技术。

申请人前两项国家自然科学基金(81000754、81471994) 研究发现：志贺菌感染及对喹诺酮耐药形势严峻，但其检测与药敏需3天以上，无法满足临床需求。表面增强拉曼光谱(SERS)技术具有快速、高灵敏度和指纹特征光谱等优势，但对临床样本分析处于探索阶段，且目前尚不能用于药敏分析。我们创新性地将该技术用于志贺菌等病原菌的快速检测及药敏分析。本课题进一步建立与优化了该新技术，并系统评估其用于临床样本中志贺菌等病原菌快速鉴定及药敏快速检测的应用价值。首先制备了适合志贺菌等病原菌分离及指纹信号增强的高性能材料，建立了标准化SERS指纹谱，构建志贺菌等病原菌指纹数据库；然后利用PEI修饰的磁性纳米颗粒分离细菌；最后利用生长的单菌落的SERS指纹谱准确鉴定志贺菌等病原菌不同血清型及耐药株。本课题的完成，为临床志贺菌等病原菌感染诊疗提供了一种快速检测的新技术与新思路。