Development of a high-throughput microfluidic integrated microarray for the detection of chimeric bioweapons.

Sheppod, Timothy ; Satterfield, Brent ; Hukari, Kyle W. ; West, Jason A. A. ; Hux, Gary A.

Sandia National Laboratories

关键词: Cloning.;    Dna-Cloning;    Classification;    Genes;    Oligonucleotides;   

DOI  :  10.2172/895702
RP-ID  :  SAND2006-5873
RP-ID  :  AC04-94AL85000
RP-ID  :  895702

美国|英语

来源: UNT Digital Library

【 摘 要 】

The advancement of DNA cloning has significantly augmented the potential threat of a focused bioweapon assault, such as a terrorist attack. With current DNA cloning techniques, toxin genes from the most dangerous (but environmentally labile) bacterial or viral organism can now be selected and inserted into robust organism to produce an infinite number of deadly chimeric bioweapons. In order to neutralize such a threat, accurate detection of the expressed toxin genes, rather than classification on strain or genealogical decent of these organisms, is critical. The development of a high-throughput microarray approach will enable the detection of unknowns chimeric bioweapons. The development of a high-throughput microarray approach will enable the detection of unknown bioweapons. We have developed a unique microfluidic approach to capture and concentrate these threat genes (mRNA's) upto a 30 fold concentration. These captured oligonucleotides can then be used to synthesize in situ oligonucleotide copies (cDNA probes) of the captured genes. An integrated microfluidic architecture will enable us to control flows of reagents, perform clean-up steps and finally elute nanoliter volumes of synthesized oligonucleotides probes. The integrated approach has enabled a process where chimeric or conventional bioweapons can rapidly be identified based on their toxic function, rather than being restricted to information that may not identify the critical nature of the threat.