DNA-based testing for environmental contamination
Inventors
Assignees
United States Department of the Army
Publication Number
US-10550392-B2
Publication Date
2020-02-04
Expiration Date
2036-09-22
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Abstract
The present invention provides synthetic DNA aptamers that bind a target explosive to allow detection of that explosive. In various embodiments, the synthetic DNA aptamers may include one or more aptamers selected from the group consisting of SEQ ID 1-6. The various synthetic DNA aptamers are sensitive to different explosives. The synthetic DNA aptamers provide an inexpensive, in situ means for testing for explosive, which may be used for both soil and water samples. This testing may include an assay method using the synthetic DNA aptamers or a biosensor linked to the synthetic DNA aptamers.
Core Innovation
This invention provides synthetic DNA aptamers that specifically bind the explosives 1,3,5-Trinitroperhydro-1,3,5-triazine (RDX) and 3-Nitro-1,2,4-triazol-5-one (NTO). These aptamers enable inexpensive, in situ methods for detecting these explosive chemicals in environmental samples such as soil and water. The invention includes assay methods that admix a buffered solution of synthetic DNA aptamers with samples for explosive testing and biosensor apparatuses that utilize these aptamers linked to electrodes to detect explosives.
The problem addressed by this invention is the current lack of rapid, in situ testing methods for RDX and NTO contamination in environmental samples. These explosives can contaminate soil and groundwater and pose health risks at low levels. Existing testing methods rely on highly sensitive, off-site instrumentation that is costly, time-consuming, and unsuitable for wide-scale environmental remediation. Additionally, current methods do not support immediate reporting or enable evaluation of contaminant dispersal patterns, creating a demand for a rapid, on-site explosive detection method.
The inventive solution involves synthetic DNA aptamers selected and optimized by systematic evolution of ligands by exponential enrichment (SELEX) to bind specifically and with high affinity to RDX or NTO. Aptamer sequences provided (SEQ ID NO. 1-6) are chemically synthesized or enzymatically produced and exhibit binding characteristics that allow formation of binding complexes with explosives. These aptamers can be employed in assays using gold nanoparticle aggregation or electrochemical biosensors that detect conformational changes upon explosive binding, allowing rapid, specific, and sensitive detection in diverse sample matrices.
Claims Coverage
The patent discloses four claims, including two independent claims directed to methods for detecting explosives using specific synthetic DNA aptamers targeting RDX and NTO.
Method for detecting RDX using synthetic DNA aptamers
A method comprising admixing a buffered solution of synthetic DNA aptamers consisting of one or more selected from SEQ ID 1-2 with a sample to be tested and assaying the sample for the explosive RDX.
Covalent linking of detectable labels to synthetic DNA aptamers for RDX detection
The method wherein the synthetic DNA aptamer is modified to covalently link to a detectable label which is covalently attached to the aptamer.
Method for detecting NTO using synthetic DNA aptamers
A method comprising admixing a buffered solution of synthetic DNA aptamers consisting of one or more selected from SEQ ID 3-6 with a sample to be tested and assaying the sample for the explosive NTO.
Covalent linking of detectable labels to synthetic DNA aptamers for NTO detection
The method wherein the synthetic DNA aptamer is modified to covalently link to a detectable label which is covalently attached to the aptamer.
The claims collectively cover methods of detecting RDX and NTO explosives by using specific synthetic DNA aptamers either unmodified or covalently linked to detectable labels, enabling selective and sensitive in situ testing.
Stated Advantages
Provides inexpensive, rapid, and in situ testing methods for explosives in soil and water.
Enables detection of explosives with high affinity and specificity using synthetic DNA aptamers.
Supports environmental remediation efforts by allowing widespread sample testing and pattern evaluation.
Facilitates immediate reporting under regulatory programs through rapid detection.
Documented Applications
In situ testing of environmental soil and water samples for contamination by the explosives RDX and NTO.
Use in environmental remediation projects to assess contamination levels and dispersal patterns over wide areas.
Deployment in biosensor apparatuses incorporating aptamer-modified electrodes for explosive detection.
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