A method for genotoxicity detection using random amplified polymorphism DNA with Danio rerio.

Abstract

The increasing presence of genotoxic pollutants in the aquatic environment has led to the development of quick monitoring methods. We have applied the random amplified polymorphism DNA (RAPD) method to evaluate the toxic effects of genotoxic chemicals. In all 22 normal wide-type Danio rerio (zebrafish), 78 amplified bands were obtained after PCR using the primer set, in which the frequency of emergence above 60% is 21. There do exist four stable bands in the amplified product, which appear in all normal test samples. The zebrafish RAPD fingerprinting database is set up on this base. The RAPD pattern from zebrafish exposed to genotoxic chemicals such as cyclophosphamide in dilution series ranging from 0.1 to 10 mg/L and dimethoate in dilution series ranging from 1.0 to 100 mg/L displayed some changes in polymorphism band patterns including the lost of stable bands. There also exists a distinct distance between the band patterns of exposed zebrafish and the control samples via the cluster method. In addition to that, the result derived from numerical analysis is more sensitive to the result obtained from stable band analysis because it can reveal the distance between the band pattern of the zebrafish exposed to 0.05 mg/L cyclophosphamide and the control sample. These results proved the accuracy and the sensitivity of the two analyses of genotoxicity effect based on RAPD fingerprinting research.