基于流式细胞术的细菌快速分型检测方法  

An efficient flow cytometry-based bacteria sorting method

作　　者：朴莹 江寅申 刘衍朋 余华海 高小炎 PIAO Ying;JIANG Yinshen;LIU Yanpeng;YU Huahai;GAO Xiaoyan(Zhejiang Key Laboratory of Smart Biomaterials,Zhejiang University,Hangzhou 310058,China;National Clinical Research Center for Child Health,Children’s Hospita,.Zhejiang University School of Medicine,Hangzhou 310012,China;Key Laboratory of Integrated Oncology and Intelligent Medicine of Zhejiang Province,Affiliated Hangzhou First People’s Hospital,Westlake University,Hangzhou 310006,China)

机构地区：[1]浙江大学浙江省智能生物材料重点实验室,浙江杭州310058 [2]浙江大学医学院附属儿童医院国家儿童健康与疾病临床医学研究中心,浙江杭州310012 [3]西湖大学附属杭州市第一人民医院浙江省肿瘤融合研究与智能医学重点实验室,浙江杭州310006

出　　处：《实验技术与管理》2023年第10期70-76,共7页Experimental Technology and Management

基　　金：浙江省公益研究计划-分析测试项目(LGC21B040001)。

摘　　要：发现一种三级胺氮氧化物聚合物胶束(OPDEA-PS)具有革兰氏阳性细菌(G+)特异性粘附且对红细胞或其他血液成分粘附相对较弱的特性。利用这一特性研究了一种基于流式细胞术的快速血液细菌分型检测方法。结果显示,将不同比例的金黄色葡萄球菌(G+)和大肠埃希菌(G–)或肺炎链球菌(G+)和铜绿假单胞菌(G–)与血液混合,加入终浓度为0.01 mg/mL的Cy5OPDEA-PS胶束,孵育10 min后用流式细胞术或平板计数法检测,得到流式检测准确率高于90%,平板计数法准确率低于80%。相比而言,流式检测法具有操作简单、快速高效、准确率高的优势。这种用流式细胞术快速分辨细菌革兰氏分型的检测方法将有助于临床上菌血症等急性血液细菌感染性疾病的细菌类型初判,达到及时诊断和治疗的目的。OPDEA-PS micelle was found to exhibit specific adhesion to Gram-positive bacteria(G+)and relatively weak adhesion to RBC(Red Blood Cell)and other blood components.A rapid blood bacterial assay by Cy5OPDEA-PS micelle based on flow cytometry was established.The results showed that after adding certain proportions of S.aureus(G+)and E.coli(G–),or S.pneumonia(G+)and P.aeruginosa(G–)with blood samples,and incubating with Cy5OPDEA-PS micelle(a final concentration of 0.01 mg/mL)for only 10 min,the accuracy of the bacteria type proportions detected and analyzed by Flow Cytometry was higher than 90%,while the accuracy of conventional colony counting method was lower than 80%.Therefore,with the advantages of simplicity,efficiency,and accuracy,this flow cytometry-based bacteria sorting method will shed light on the early detection and diagnosis of bacteremia and other acute hematological infectious diseases.

关 键 词：流式细胞术 细菌 血液感染 快速检测 

分 类 号：R-331[医药卫生]