Accession Number : ADA259050


Title :   Rapid Detection of Infectious Agents with a Biosensor Based Nucleic Acid Hybridization Assay


Descriptive Note : Final rept. 30 Sep 1991-30 Sep 1992


Corporate Author : MOLECULAR DEVICES CORP MENLO PARK CA


Personal Author(s) : Hahnenberger, Karen M


Full Text : https://apps.dtic.mil/dtic/tr/fulltext/u2/a259050.pdf


Report Date : 18 Dec 1992


Pagination or Media Count : 28


Abstract : The goal of this project has been to optimize the Threshold DNA probe assay as a method for quantitating specific sequences after amplification by the polymerase chain reaction. Increasing the probe concentration 5-fold in the standard scheme B assay results in more rapid annealing of the probes to the target DNA, decreasing the overall assay time to less than 1 hour. A second assay format was developed in which the capture reagent and enzyme conjugate are added simultaneously. This scheme A format gives comparable results to those obtained using scheme B, and shortens the assay time to less than 30 minutes. Studies were also initiated to increase the detection sensitivity of the Threshold DNA probe assay by introducing multiple labels into the probe sequence by PCR. Data obtained after the reporting period officially ended indicate that 10 genomic targets can be detected after 25 cycles of PCR by using longer probes containing multiple fluorescein labels.


Descriptors :   *DETECTION , *SENSITIVITY , *DEOXYRIBONUCLEIC ACIDS , PROBES , ENZYMES , REDUCTION , STANDARDS , CHAIN REACTIONS , HYBRIDIZATION , NUCLEIC ACIDS , AMPLIFICATION , FORMATS , SEQUENCES , CYCLES , TARGETS , ANNEALING


Subject Categories : Medicine and Medical Research
      Biomedical Instrumentation and Bioengineering


Distribution Statement : APPROVED FOR PUBLIC RELEASE