量子点-磁微粒技术同步检测输血后肝炎的体系建立及初步应用

Establishment and application of the synchronous detection system of post-transfusion hepatitis based on quantum dots and magnetic microspheres technology

目的结合量子点-磁微粒(QD-MMS)技术,建立针对HBV及HCV的多抗原同步快速检测体系。方法以碳二亚胺交联法活化QD-MMS以夹心法检测抗原。在分别建立HBV与HCV抗原检测方法的基础上整合优化体系,对188份随机临床标本进行同步检测,结果与聚合酶链反应(PCR)及酶联免疫吸附法(ELISA)方法比较。结果 QD-MMS同步检测体系检测范围为HbsAb 2ng/mL～10μg/mL、HCcAg 5ng/mL～5μg/mL,40min内完成同步检测。与ELISA方法比较灵敏度及特异性无差异,符合率分别为95.8%、98.4%。与TP、HAV阳性标本无交叉反应。试剂于4℃保存4周后使用,与新配制试剂检测结果无差异。结合目标抗原后的复合物标本,经4℃放置4周后荧光强度无明显衰减。最长放置8周后仍可检出荧光。结论建立了对HBsAg和HCcAg的同步检测体系,采用图像分析软件解析荧光强度值可达到半定量检测效果。检测时间同步缩短到了40min左右,提高了检测效率,操作简便快速,检测成本低,所需样本量少,特异度较好,能初步满足多抗原同步检测的需求,为同步检测更多病原体抗原体系的建立奠定了基础。

Objective To establish a synchronous detection system against multiple antigens of HBV and HCV based quantum dots（QDs） and magnetic microspheres（MMS）.Methods Sandwich method was constructed with QDs and MMS activated with carbodiimide cross-link technique.On the basis of single test systems,the optimal detection system was constructed and synchronously performed to detect 188 unknown samples.The detection results were compared with those of polymerase chain reaction（PCR） and enzyme linked immunosorbent assay（ELISA）.Results The detecting ranges were 2 ng/mL-10 μg/mL for HBsAb and 5 ng/mL-5 μg/mL for HCcAg.The time of detection were shortened in 40 min.And there were no significant difference between ELISA and this method,with the accordance rates of 95.8% and 98.4% for HBsAb and HCcAg in parallel test system.The fluorescence intensity of compound samples did not decline,after being stored at 4 ℃ for 4 weeks.The longest storing period could be 8 weeks.Conclusion The synchronous detection system of HBsAg and HCcAg and semi-quantitative detection could be achieved by using image analyzing software.The measure period could be shortened to 40 min in parallel,with advantages of high detection efficiency,fast detection speed,low cost,low requirement of sample volume and high specificity,could be fit for synchronous detection of multiple antigens.This technology might be helpful for parallel detection of antigens.