免疫磁性分离技术在病原微生物检测中的应用

doi:10.16431/j.cnki.1671-7236.2024.05.042

摘要/Abstract

摘要： 免疫磁性分离技术是通过磁场作用使得磁珠表面所吸附的靶标物从样品中分离的前处理技术。免疫磁珠捕获靶标后,免疫磁珠-靶标复合物磁吸固定在管壁,洗脱杂质即可得到靶标。免疫磁性分离技术富集效果主要受抗体特异性、磁珠直径、磁珠添加量、孵育时间和样本pH影响,其中抗体特异性是影响磁珠富集效果的关键。利用免疫磁珠可吸附抗体的特性,在酶联免疫吸附测定方法中替代包被步骤,可大幅减少检测时间;免疫磁性分离技术作为前处理方法,与其他检测方法联用后,可提高检测方法的灵敏度和检测效率。笔者从免疫磁性分离技术原理、一般分离步骤、常见影响因素,以及在酶联免疫吸附测定、免疫层析法、PCR、实时荧光定量PCR、环介导等温扩增反应、流式细胞术、电化学传感器、液相色谱法等应用方面进行阐述,为免疫磁性分离技术的研究及推广提供参考。

关键词: 免疫磁性分离技术; 样本前处理; 病原微生物

Abstract: Immunomagnetic separation technology is a pre-treatment technology that separates the target adsorbed on the surface of the magnetic bead from the sample under the magnetic field.After the immune magnetic bead captures the target,the immunemagnetic bead-target complex is magnetically fixed on the tube wall,and the target can be obtained by elution of impurities.The enrichment effect of immunomagnetic separation technology is mainly affected by antibody specificity,magnetic bead diameter,magnetic bead addition amount,incubation time and sample pH,in which antibody specificity is the key to affect the enrichment effect of magnetic beads.Using the characteristics of immunomagnetic beads to adsorb antibodies,the coating step can be replaced in the enzyme-linked immunosorbent assay method,which can greatly reduce the detection time.As a pre-treatment method,immunomagnetic separation technology can improve the sensitivity and efficiency of the detection method when combined with other detection methods.In this review,the technical principle,general separation steps,common influencing factors and the application in enzyme-linked immunosorbent assay,immunochromatography,PCR,Real-time quantitative PCR,loop-mediated isothermal amplification reaction,flow cytometry,electrochemical sensors,liquid chromatography were described,so as to provide reference for the research and application of immunomagnetic separation technology.

Key words: immunomagnetic separation technology; sample preparation; pathogenic microorganism

中图分类号:

张峻豪, 匡磊, 雷镒妃, 赵天睿, 许灏钧, 陈斌, 王丹, 韦阳飞, 洪灯, 胡长敏. 免疫磁性分离技术在病原微生物检测中的应用[J]. 中国畜牧兽医, 2024, 51(5): 2219-2227.

ZHANG Junhao, KUANG Lei, LEI Yifei, ZHAO Tianrui, XU Haojun, CHEN Bin, WANG Dan, WEI Yangfei, HONG Deng, HU Changmin. Application of Immunomagnetic Separation Technology in the Detection of Pathogenic Microorganisms[J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(5): 2219-2227.

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