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检测致癌物-DNA加合物的免疫学方法。

Immunological methods for detection of carcinogen-DNA adducts.

作者信息

Adamkiewicz J, Nehls P, Rajewsky M F

出版信息

IARC Sci Publ. 1984(59):199-215.

PMID:6085833
Abstract

Considerable advances have been made during recent years, with regard to the detection and quantification of carcinogen- or mutagen-induced, structural modifications in the DNA of mammalian cells, by the introduction of immunoanalytical methods, in particular in conjunction with monoclonal antibodies (Mab). Antibodies are characterized by an outstanding capacity for the specific recognition of subtle alterations of molecular structure. They can, therefore, be used as sensitive detection probes in assays for DNA modifications caused by low levels of DNA-reactive (e.g., environmental) agents. Depending on the purpose of analysis, various types of immunoassays can be performed. The competitive radioimmunoassay (RIA) represents a routinely applicable, reproducible and sensitive assay for the detection of defined carcinogen-DNA adducts in hydrolysates of cellular DNA, in body fluids or in urine. Depending on their particular design, enzyme immunoassay (EIA) may have exceptionally low detection limits, due to the enzymatic amplification of the measured radioactivity or colour intensity. Similarly, recently established immuno-slot-blot (ISB) techniques are also characterized by very high sensitivity. Immunocytological assays (ICA) use Mab in conjunction with electronically intensified immunofluorescence for detection of modified DNA components in the nuclei of individual cells. Finally, single modified deoxynucleosides can be detected and localized in individual DNA molecules by immuno-electron microscopy (IEM).

摘要

近年来,在通过引入免疫分析方法,特别是与单克隆抗体(Mab)结合来检测和定量哺乳动物细胞DNA中致癌物或诱变剂诱导的结构修饰方面取得了相当大的进展。抗体的特点是具有特异性识别分子结构细微变化的卓越能力。因此,它们可以用作检测由低水平DNA反应性(如环境)剂引起的DNA修饰的灵敏检测探针。根据分析目的,可以进行各种类型的免疫测定。竞争性放射免疫测定(RIA)是一种常规适用且可重复的灵敏测定方法,用于检测细胞DNA水解产物、体液或尿液中特定的致癌物-DNA加合物。根据其特定设计,酶免疫测定(EIA)可能具有极低的检测限,这是由于测量的放射性或颜色强度的酶促放大作用。同样,最近建立的免疫斑点印迹(ISB)技术也具有非常高的灵敏度。免疫细胞分析(ICA)将Mab与电子增强免疫荧光结合使用,以检测单个细胞核中修饰的DNA成分。最后,可以通过免疫电子显微镜(IEM)在单个DNA分子中检测和定位单个修饰的脱氧核苷。

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