Rapid, potentially automatable, method extract biomarkers for HPLC/ESI/MS/MS to detect and identify BW agents

Conference · Wed Dec 31 04:00:00 EST 1997

OSTI ID:661538

White, D C ^[1]; Burkhalter, R S; Smith, C ^[1]; Whitaker, K W ^[2]

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1. Univ. of Tennessee, Knoxville, TN (United States). Center for Environmental Biotechnology
2. Microbial Insights, Inc., Rockford, TN (United States)

The program proposes to concentrate on the rapid recovery of signature biomarkers based on automated high-pressure, high-temperature solvent extraction (ASE) and/or supercritical fluid extraction (SFE) to produce lipids, nucleic acids and proteins sequentially concentrated and purified in minutes with yields especially from microeukaryotes, Gram-positive bacteria and spores. Lipids are extracted in higher proportions greater than classical one-phase, room temperature solvent extraction without major changes in lipid composition. High performance liquid chromatography (HPLC) with or without derivatization, electrospray ionization (ESI) and highly specific detection by mass spectrometry (MS) particularly with (MS){sup n} provides the detection, identification and because the signature lipid biomarkers are both phenotypic as well as genotypic biomarkers, insights into potential infectivity of BW agents. Feasibility has been demonstrated with detection, identification, and determination of infectious potential of Cryptosporidium parvum at the sensitivity of a single oocyst (which is unculturable in vitro) and accurate identification and prediction, pathogenicity, and drug-resistance of Mycobacteria spp.

• Research Organization: Oak Ridge National Lab., TN (United States)
• Sponsoring Organization: USDOE Assistant Secretary for Human Resources and Administration, Washington, DC (United States)
• DOE Contract Number: AC05-96OR22464
• OSTI ID: 661538
• Report Number(s): ORNL/CP--97757; CONF-971163--; ON: DE98003517
• Country of Publication: United States
• Language: English

Related Subjects

45 MILITARY TECHNOLOGY, WEAPONRY, AND NATIONAL DEFENSE
BACTERIA
BIOLOGICAL MARKERS
BIOLOGICAL WARFARE AGENTS
GENOTYPE
INFECTIVITY
LIQUID COLUMN CHROMATOGRAPHY
MASS SPECTROSCOPY
PHENOTYPE