摘要
目的探讨免疫比浊结合免疫磁珠分离技术在嗜麦芽窄食单孢菌自动快速定量检测中的初步应用。方法嗜麦芽窄食单孢菌免疫日本大耳白兔制备多抗血清。ProteinA磁珠与多抗偶联制成多抗免疫磁珠,以该磁珠捕获模拟标本的嗜麦芽窄食单孢菌,接种培养后菌落计数,计算免疫磁珠的特异富集作用。富含SPA的金黄色葡萄球菌(No1800株)经甲醛灭活后,与嗜麦芽窄食单孢菌的多抗血清结合,制备特异抗体致敏的金黄色葡萄球菌,作为协同凝集试剂和自动定量检测的免疫比浊试剂,在日立7060全自动生化分析仪编制程序进行自动检测和初步的方法学评价。结果嗜麦芽窄食单孢菌免疫日本大耳白兔获得的多抗血清特异性好,凝集效价达1∶240;制备的多抗免疫磁珠对嗜麦芽窄食单孢菌的捕获效率约为35%;抗体致敏的金黄色葡萄球菌能与待测嗜麦芽窄食单孢菌产生明显的凝集现象,在日立7060全自动生化分析仪上定量检测其灵敏度好、线性范围宽、特异性良好,检测结果与SYSMEXUF-50全自动尿液分析仪基本一致。结论制备特异的多抗磁珠对嗜麦芽窄食单孢菌有一定的特异捕获与富集作用,协同凝集试剂与免疫比浊试剂对嗜麦芽窄食单孢菌的自动、快速、定量检测具有较好的效果,为其病原菌的快速诊断和自动定量分析提供了新思路。
Objective To establish a nephelometric immunoassay for the rapid automatic detection of Stenotrophomonas maltophilia. Methods The specific antiserum was prepared by rabbits immunized with Stenotrophomonas maltophilia and the magnetic beads coated with SPA binding to the specific IgG from the antiserum were used to capture the Stenotrophomonas maltophilia from the samples..SPA-rich Staphylococcus aureus killed by formalin was also combined with the specific IgG from the antiserum and then used as a nephelometric immunoassay reagent.The rapid automatic quantitative detection of Stenotrophomonas maltophilia was performed on the Hitachi 7060 biochemical analyzer. Results In the present study,the high quality antiserum with agglutination titer of 1:240 and high specificity of Stenotrophomonas maltophilia was got from the immunized rabbits.Immunomagnetic beads had some enrichment effects of Stenotrophomonas maltophilia,and the colony counting of plate culture was performed after enriched by immunomagnetic beads,indicating that the capture efficiency was about 35%.The rapid automatic quantitative detection method of Stenotrophomonas maltophilia was established on the Hitachi 7060 analyzer with the SPA-rich Staphylococcus aureus combined with the specific IgG used as the nephelometric immunoassay reagent.This immunoassay has a good sensitivity and relatively satisfied specificity and linear range. Conclusion With the immunomagnetic beads and the SPA-rich Staphylococcus aureus combined with the specific IgG of the antiserum,the rapid automatic quantitative immunoassay for the detection of Stenotrophomonas maltophilia is well established,which provides a new way for the diagnosis of pathogenic bacteria.
出处
《总装备部医学学报》
2010年第4期198-201,共4页
Medical Journal of General Equipment Headquarters
关键词
嗜麦芽窄食单孢菌
免疫磁珠
免疫比浊
快速定量分析
Stenotrophomonas maltophilia
immunomagnetic
nephelometric immunoassay
rapid automatic quantitative analysis of bacteria