30 November 1999 Nonenzymatic microorganism identification based on ribosomal RNA
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Effective defense against biological warfare (BW) agents requires rapid, fieldable and accurate systems. For micro- organisms like bacteria and viruses, ribosomal RNA (rRNA) provides a valuable target with multiple advantages of species specificity and intrinsic target amplification. Vegetative and spore forms of bacteria contain approximately 104 copies of rRNA. Direct detection of rRNA copies can eliminate some of the interference and preparation difficulties involved in enzymatic amplification methods. In order to apply the advantages of rRNA to BW defense, we are developing a fieldable system based on 16S rRNA, physical disruption of the micro-organism, solid phase hybridization, and fluorescence detection. Our goals include species-specific identification, complete operation from raw sample to identification in 15 minutes or less, and compact, fieldable instrumentation. Initial work on this project has investigated the lysis and hybridization steps, the species-specificity of oligonucleotides probes, and the development of a novel electromagnetic method to physically disrupt the micro- organisms. Target bacteria have been Escherichia coli (E. coli) and Bacillus subtilis (B. subtilis). Continuing work includes further development of methods to rapidly disrupt the micro-organisms and release the rRNA, improved integration and processing, and extension to bacterial and mammalian viruses like MS2 and vesicular stomatitis virus.
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Jeffrey T. Ives, Jeffrey T. Ives, Alicia M. Pierini, Alicia M. Pierini, Jeffrey A. Stokes, Jeffrey A. Stokes, Thomas M. Wahlund, Thomas M. Wahlund, Betsy Read, Betsy Read, James H. Bechtel, James H. Bechtel, Burt V. Bronk, Burt V. Bronk, "Nonenzymatic microorganism identification based on ribosomal RNA", Proc. SPIE 3855, Air Monitoring and Detection of Chemical and Biological Agents II, (30 November 1999); doi: 10.1117/12.371268; https://doi.org/10.1117/12.371268

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