Fiscal Year:
2009
Title:
Rapid Microbial Identification by MALDI-TOF Mass Spectrometry of Ribosomal RNA
Agency:
HHS
Contract:
2R44AI066425-02
Award Amount:
$722,491.00
Abstract:
DESCRIPTION (provided by applicant): The World Health Organization estimates that over 1,600 people die each hour from an infectious disease, half of whom are children under 5 years of age. Infectious diseases account for 26 percent of total global mortali
ty and are the third leading cause of death in the United States. In addition to microbial identification for clinical response, diagnostics are also needed for a wide range of applications including food and water safety, bioreactor analysis, sterility as
surance in the pharmaceutical industry, environmental microbiology, and homeland security. The focus of this research is to develop and characterize a microbial identification platform using very rapid MALDI-TOF mass spectrometry of modified ribosomal RNA
fragments. By generating base-specific RNA fragments optimized for mass spectrometric characterization, we have shown that organism-pure samples such as single colonies and clones can be identified in ca. 1.5 hours. The platform is open in that it can
identify any bacterial organism that has been previously sequenced without the a priori design of sequence specific nucleic acid probes. Furthermore, the assay is more rapid than nucleic acid sequencing and amenable to identification of organisms in biolog
ical mixtures and determination of their relative abundances. Finally, pathogens can be identified against an uninteresting background of commensal organisms or host nucleic acids. We plan to build on our promising system for general bacterial identificati
on by improving our techniques for nucleic acid fractionation based upon relative organism abundance, heuristic approaches, and clonal PCR for organism-mixture analysis. The anticipated result of this study will be a rapid, first response system capable of
high-throughput microbial community analysis of a variety of sample types. Including nucleic acid isolation, data acquisition and analysis, ca. 384 analyses should be feasible in less about 5 hours. PUBLIC HEALTH RELEVANCE: Rapid response to infectious di
sease requires that the analytical procedures employed are inherently fast. Compared to most analytical techniques, mass spectrometry is generally faster, providing both separation of complex mixtures and a fundamental measurement simultaneously. In additi
on to microbial identification for clinical response, diagnostics are also needed for a wide range of applications including food and water safety, bioreactor analysis and sterility assurance, and environmental microbiology. This project focuses on develop
ing a total platform system for microbial identification using predictive software, novel molecular techniques, and mass spectral analysis of ribosomal RNA fragments.
Small Business Information at Submission:
BIOTEX, INC.
BIOTEX, INC. 8058 EL RIO ST HOUSTON, TX 77054
EIN/Tax ID:
742824041
DUNS:
N/A
Number of Employees:
N/A
Woman-Owned:
No
Minority-Owned:
No
HUBZone-Owned:
No