量子点荧光探针流式技术定量检测血清降钙素原的方法学及临床应用  

Establishment of a quantum-dot cytometric assay for detecting PCT and the value of clinical detection

作　　者：胡晓璐[1] 王浈[2] 祝仲珍[2] 王占科[2] 

机构地区：[1]南昌大学医学院研究生院,江西南昌330006 [2]解放军第94医院检验科

出　　处：《中国老年学杂志》2013年第8期1828-1831,共4页Chinese Journal of Gerontology

基　　金：国家科技支撑项目(重大疾病综合检测生物芯片研究与开发)(2008BAI52B03);江西省科技支撑项目(2008BA07400)

摘　　要：目的将量子点探针与流式技术结合,建立定量检测血清降钙素原(PCT)的新方法并对检测性能及临床应用进行评价。方法将羊抗PCT抗体与微球偶联制备诊断微球,与血清或标准品中的PCT结合,加入鼠抗PCT抗体,最后加入生物素化的羊抗鼠IgG抗体和链霉亲和素化量子点,流式细胞仪检测平均荧光强度(MFI)。选择2012年1月至2012年8月入住本院重症监护病房(ICU)77例老年创伤患者,通过本实验创建的定量检测PCT方法对77例老年创伤患者入院第1、3、5、7、10 d进行血清PCT检测,并记录当日急性生理学与慢性健康状况评分系统(APACHEⅡ)、C-反应蛋白(CRP)等,比较PCT水平、APACHEⅡ评分及CRP水平,分析各指标之间的相关性。结果自创法检测PCT的灵敏度为5.42 pg/ml、线性范围为45.42 pg/ml～10 ng/ml,针对老年群体阳性下限值为0.56 ng/ml。采用自创法检测PCT水平预测老年创伤患者发生感染的灵敏度为100%,特异度为24.5%,准确度为84.4%,PCT水平监测老年创伤病情和感染预测方面优于CRP且与APACHEⅡ评分相关性好。结论本实验创建的定量检测血清PCT的方法可作为临床检测血清PCT的方法,早期诊断及监测老年创伤患者合并感染的灵敏度、特异度较高,适用于临床诊断。Objective To establish a quantum dots cytometric assay for detecting PCT and studying its precision, linearity, sensitiv-ity. Methods Microsphere surface was labeled by sleep against PCT antibody, combined with PCT, adding rabbit against PCT antibody,last added biofinylation sleep against rabbit IgG antibody and QDs-SA conjugate. The mean fluorescent intensity （MFI） was detected by flow cy-tometer. Elderly-patients with heavy-hurt enrolled in ICU were retrospectively investigated and analyzed, using the new assay for detecting PCT. Results The sensitivity（45.42 pg/ml）, linearity（45.42 pg/ml - 10 ng/ml） of quantum-dots cytometric assay were better. The posi-tive limit was 0. 56 ng/ml. A method to detect PCT prediction elderly trauma in patients with the sensitivity of infection was 100%, respec-tively was 24. 5% ,the accuracy was 84.4%, PCT had a good correlation with APACHE Ⅱ. Conclusions The method could diagnose and monitor old trauma patients merge infection with higher sensitivity and specific degrees, which is suitable for clinical diagnosis.

关 键 词：量子点 流式技术 降钙素原 平均荧光强度 

分 类 号：R631[医药卫生—外科学]