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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Carrillo, Maria P. Bruno, John G. Richarte, Alicia M. Edge, Allison |
| Description | Country affiliation: United States Author Affiliation: Bruno JG ( Operational Technologies Corporation, NW Loop 410, Suite 230, San Antonio, TX 78229, USA. john.bruno@otcorp.com) |
| Abstract | Sixty candidate DNA aptamers were developed against botulinum neurotoxin (BoNT) type A light chain (LC) from ten rounds of selection, resulting in several identical sequences. Secondary structures of the identical aptamers were compared to structures of previously reported BoNT A DNA aptamers. A series of ten candidate loop structures were selected from this comparison as potential binding pockets and aptamer beacons. These candidate beacons were synthesized with 5'-TYE 665 and 3'-Iowa Black quencher labels for comparison of fluorescence levels as a function of BoNT A LC concentration. Only three of the ten candidates exhibited any fluorescence response to increasing levels of BoNT A LC. However, of the two most responsive candidates, one represented a subset loop of the larger more intensely fluorescent double-looped structure, designated Beacon 10. This beacon yielded a lower limit of detection of 1 ng/mL in buffer using a spectrofluorometer and a portable handheld fluorometer, but also responded substantially to BoNT A, B, E holotoxins and heavy or light chain components even in a dilute soil suspension, but not in 50% human serum. Beacon 10 did not respond strongly to a variety of other divergent peptides, suggesting that it is relatively specific to the level of botulinum toxins and is only useful for environmental testing. Beacon 10 also shared short sequence segments with other published BoNT aptamer DNA sequences, suggesting that these may be points of physical contact between the aptamers and BoNTs. |
| ISSN | 09565663 |
| Issue Number | 1 |
| Volume Number | 31 |
| e-ISSN | 18734235 |
| Journal | Biosensors and Bioelectronics |
| Language | English |
| Publisher | Elsevier |
| Publisher Date | 2012-01-15 |
| Publisher Place | Great Britain (UK) |
| Access Restriction | One Nation One Subscription (ONOS) |
| Subject Keyword | Aptamers, Nucleotide Chemistry Biosensing Techniques Instrumentation Botulinum Toxins Analysis Molecular Probe Techniques Genetics Equipment Design Equipment Failure Analysis Reproducibility Of Results Sensitivity And Specificity Journal Article Research Support, N.i.h., Extramural Research Support, U.s. Gov't, Non-p.h.s. Discipline Biotechnology |
| Content Type | Text |
| Resource Type | Article |
| Subject | Nanoscience and Nanotechnology Medicine Biophysics Biomedical Engineering Biotechnology Electrochemistry |
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