人肺炎支原体抗原量子点免疫层析法的建立

Detection of human Mycoplasma pneumoniae antigen by double antibodies sandwich method based on quantum dots immunochromatography

目的采用量子点标记技术和免疫层析技术,建立人肺炎支原体抗原的检测体系,并对其临床应用进行初步探讨。方法在偶联剂碳二亚胺EDC和N-羟基琥珀酰亚胺(N-hydroxysuccinimide,NHS)作用下,使兔抗人肺炎支原体(Mp)多克隆抗体与羧基量子点(Cd Se/Ze S)偶联,制备量子点标记物,以针对Mp P1蛋白的单抗作为捕获抗体,建立双抗体夹心量子点免疫层析法检测肺炎支抗原。该方法检测50份临床咽拭子标本,并与培养法相比较。结果量子点免疫层析法检测临床标本Mp,阴性符合率为96.6%;阳性符合率为100%;最低检出浓度为0.5 ng/m L。结论量子点(Cd Se/Zn S)标记免疫层析法检测肺炎支原体与Mp培养法具有较好的符合性,该方法操作简便、检测快速、灵敏度高,适用于Mp感染的早期诊断。

Objective To establish a double antibody-sandwich method based on quantum dots immunochromatographyfor detection of human Mycoplasma pneumoniae antigen. Methods Rabbit anti – Mycoplasma pneumoniae polycloneantibody was labeled with coupling agent, Cd Se/Ze S quantum dots（QDs） by using covalent bonding of EDC and NHS andquantum dots immunochromatography was established with monoclonal antibody to Mp Plus capture antibody and this methodwas used for detection of 50 copies of clinical throat swabs. The results were compared with culture method. Results Thenegative coincidence rate of quantum dots immunochromatography in detection of 50 copies of throat swabs were 96.6% andthat of positive coincidence rate was 100%. The minimum detectable concentration was 0.5ng/m L. Conclusion Quantumdots immunochromatography is easy to perform with high sensitivity and it is suitable for early and rapid diagnosis of Mycoplsma pneumpniae infection.