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Title: Novel Mass Spectrometry Mutation Screening for Contaminant Impact Analysis

Due to the limited budget of waste clean-up for all DOE contamination sites, it is critical to have a sound risk analysis with strong scientific basis to set priority for waste clean-up. In the past, the priority was often determined mostly by the type and quantity of pollutants and the observation of cancer rate increase. Since human cancers can be caused by various reasons in addition to environmental contamination, a rigorous study to find the relationship between specific contaminants and cancer is critically important for setting up the priority for waste clean-up. In addition, a contaminated site usually contain many different pollutants. However, it can be only a few specific pollutants are carcinogenic chemicals which are responsible for most cancers. Clean-up of small quantity of critical pollutants instead of the entire pollutant site can save significant decontamination cost. Since a few anti-tumor genes such as p53 and ras genes are highly conserved among various animals and mutation of these genes have been associated with many human cancers, it is very valuable to find the relationship between specific contaminant and specific cancer. Since it is not possible to pursue any human on the relationship of cancer and specific pollutant under wellmore » defined experimental conditions, it is desirable to pursue experiments on animals such as fish and mice to find out the relationship of mutation of p53 gene and specific contaminant. It is also required that the sequence of the region of p53 gene in animal is same as human being. Mutations due to pollutant can happen at various sites and only occur at a small percentage. In order to confirm the relationship between specific pollutant and mutation, a very large number of DNA samples need to be carefully analyzed. In the past, nearly all DNA analyses were pursued by gel electrophoresis. It is relatively slow and expensive. It is not feasible to obtain the relationship of mutations with specific contaminant with present DNA analysis technology. Thus, our approach is to develop novel new DNA technologies which can potentially achieve rapid, reliable and inexpensive DNA analysis for environmental applications. The objective of this program is to develop innovative mass spectrometry technology to achieve fast mutation screening and to reveal the linkage between gene mutation and contaminants. Mass spectrometry has the potential to achieve very fast speed sample analysis.New innovative approaches for improving mass resolution and detection sensitivity were pursued to help to achieve rapid DNA screening. Allele specific polymerase chain reaction (ASPCR) coupled with mass spectrometry for DNA mutation detection was also pursued. This technology was applied to wildlife species such as fish for the genotoxic effect of hazardous waste to be assessed at DNA level.« less
Authors:
;
Publication Date:
OSTI Identifier:
829893
Report Number(s):
EMSP-60218
R&D Project: EMSP 60218; TRN: US200429%%66
Resource Type:
Technical Report
Resource Relation:
Other Information: PBD: 30 Sep 2000
Research Org:
Oak Ridge National Lab., Oak Ridge, TN (US)
Sponsoring Org:
USDOE Office of Environmental Management (EM) (US)
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; 54 ENVIRONMENTAL SCIENCES; CONTAMINATION; DECONTAMINATION; DETECTION; DNA; ELECTROPHORESIS; GENE MUTATIONS; GENES; MASS RESOLUTION; MASS SPECTROSCOPY; MICE; MUTATIONS; NEOPLASMS; POLLUTANTS; POLYMERASE CHAIN REACTION; RISK ASSESSMENT; VELOCITY; WASTES