DNA指纹技术在污染土壤生态毒理诊断中的应用

Application of DNA fingerprint in eco-toxicological diagnosis of contaminated soil.

生物标记物能在细胞或分子水平上指示暴露-效应关系,是进行污染土壤生态毒理诊断的主要技术手段之一。随着分子生物学技术的飞速发展,出现了一系列以聚合酶链式反应为基础的、在分子水平上检测污染物质导致的生物体DNA损伤的DNA指纹技术。DNA指纹技术的主要类型有:随机扩增多态性DNA(RAPD)、聚合酶链式反应-单链构象多态性(PCR-SSCP)、扩增片段长度多态性(AFLP)、任意引物聚合酶链式反应(AP-PCR)、差异显示反转录聚合酶链式反应(DDRT)、短DNA重复序列(SSR)及限制片段长度多态性(RFLP)等。这些技术与检测基因突变、染色体畸变和损伤为主的一系列经典研究方法如彗星分析、微核实验等相比具有简便、快速、灵敏等优点。本文着重介绍了随机扩增多态性DNA、聚合酶链式反应-单链构象多态性、扩增片段长度多态性3种重要的DNA指纹技术在污染土壤诊断中的应用。

Biomarker can show the exposure-effect relationships at molecular and cellular levels, and is one of the most effective techniques in eco-toxicological diagnosis of contaminated soil. With the progress of molecular biology, a number of selective and sensitive PCR-based DNA fingerprint assays were developed to detect the DNA lesions caused by pollutants. The DNA fingerprint techniques mainly include random amplified polymorphic DNA （RAPD）, polymerase chain reaction-single strand conformation polymorphism, amplified fragment length polymorphism, arbitrary primed-polymerase chain reaction, differential display reverse transcription, short-sequence DNA repeat, and restriction fragment length polymorphism. These techniques possess the characters of simplicity, reproducibility and sensitivity, compared with classical methods such as comet assay and micronucleus test. In this paper, the application of three main DNA fingerprints in eco-toxicological diagnosis of contaminated soil was discussed emphatically.