阵列微流控技术在血感染病原微生物快速培养、分离和鉴定中的应用

项目名称： 阵列微流控技术在血感染病原微生物快速培养、分离和鉴定中的应用

项目编号： No.31470221

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 微生物学

项目作者： 杜文斌

作者单位： 中国科学院微生物研究所

项目金额： 82万元

中文摘要： 败血症的快速诊断是一个世界性的难题。本课题将从加快培养速度、实现血液致病菌的直接高效富集、实现高灵敏高特异性检测和耐药性筛查四个方面进行系统深入的研究。1）通过保证血培养样品的总体积在毫升级，将样品等分分配到微升体积的培养单元中，以加快培养速度，并通过选择性过滤和趋化作用实现血细胞和微生物的分隔和对早期生长的监测；2）通过微生物和血细胞在介电极化特性的差异，实现广谱高效的微生物介电电泳选择性捕获，为血液微生物直接检测和培养后检测提供有效的低浓度细菌分离富集方法；3）利用微流控芯片实现富集病原体阵列基因扩增检测和芯片原位基质辅助解析质谱指纹图谱鉴定；4）建立阵列式耐药性测试方法，快速测定微生物的耐药类型和耐药浓度。基于以上功能单元的开发与集成，有望实现低成本、高灵敏的快速血液感染广谱鉴定，缩短败血症诊断周期。以上方法也可应用于其他感染疾病诊断、食品安全检测和环境微生物群落分析等。

中文关键词： 病原微生物；败血症；快速诊断；微流控

英文摘要： Rapid diagnosis of sepsis is still an unresolved problem of the modern era. In this proposal, we will carry out research work in four aspects including accelerating blood culture, realizing fast enrichment of pathogens in whole human blood, providing high sensitivity and high specificity on-chip detection, and rapid drug resistance screening. First, we will introduce an array culture method with small unit volume to increase the initial pathogen concentration and accelerate the growth, we will introduce a simple way to selectively filtrate blood cells and allow early growth of pathogens to be detected by imaging; second, we will develop and validate a broad-range separation approach to efficiently separate and enrich pathogen cells by dielectrophoresis; third, on-chip detection of captured pathogens will be realized by multiplex PCR and MALDI-TOF mass spectrometry; finally, we will introduce a chip-based antimicrobial susceptibility test for rapid determination of the causative pathogens. We envision that the methods developed in this research can provide a practical solution for early sepsis diagnosis, and it can be adopted for other applications such as food safety testing, water quality analysis and assessment of microbial communities in environment.

英文关键词： pathogen;sepsis;rapid diagnostics;microfludics