非荧光法与荧光法芯片检测细菌感染的研究

Rapid detection of infection with bacteria by fluorescence and non-fluorescence microarray system

目的 研制一种用于快速检测急症常见感染细菌的芯片反应系统。方法 应用生物信息学技术,设计检测细菌的探针和聚合酶链反应扩增引物,探针点制成寡核苷酸芯片,待测样本经扩增并标记生物素或CY5荧光素后与芯片杂交,再经链酶亲和素碱性磷酸酶显色反应后,获得肉眼可见的杂交信号,或通过荧光检测仪读出样本的检测结果。结果 建立了针对临床急症患者标本的非荧光法与荧光法芯片检测系统,芯片的检测灵敏度(大肠埃希菌)为10～10 0CFU/反应体系。结论 两种方法不仅能快速、灵敏地检测靶细菌感染,且重复性好、信号强及不易出现非特异信号。生物素酶联显色芯片法由于更为经济、简便而有重要的临床价值。

Objective To develop a rapid detection system for clinical pathogenic bacteria. Methods Probes and primers were designed according to the sequence of pathogenic bacteria, and a microarray-based detection system was developed. DNA of standard bacteria strains was amplified and labelled by biotin or CY5, and hybridized with the micro array. Streptavidine-AP was applied in chromotest or laser scanner was used in fluorescence array. Results A microarray-based bacterial detection system was successfully developed for clinical use. Conclusion Two methods can both detect infection rapidly, sensitively and specifically. Biotin, streptavidine-AP coloration DNA detection chip has important clinical value for its economic cost and simplicity.