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A PCR Jet for Rapid Detection of Special pathogens

Award Information

Agency:
Department of Health and Human Services
Branch:
N/A
Award ID:
60507
Program Year/Program:
2004 / SBIR
Agency Tracking Number:
AI052936
Solicitation Year:
N/A
Solicitation Topic Code:
N/A
Solicitation Number:
N/A
Small Business Information
Megabase Research Products
4711 Huntington Ave., Suite 2W Lincoln, NE 68504
View profile »
Woman-Owned: Yes
Minority-Owned: No
HUBZone-Owned: No
 
Phase 2
Fiscal Year: 2004
Title: A PCR Jet for Rapid Detection of Special pathogens
Agency: HHS
Contract: 2R44AI052936-02
Award Amount: $743,361.00
 

Abstract:

DESCRIPTION (provided by applicant): Polymerase Chain Reaction (PCR) amplification of DNA is the most reliable method for the rapid detection of pathogens. The overall rate of the PCR process is determined by performance of both the thermocycler and the companion biochemistry used to enzymatically copy DNA. In Phase I Research, a very fast experimental PCRJet thermocycler has been fabricated. This pressurized gas machine with electronic valves can carry out 30 PCR cycles with thermal control + 0.2 xC in <1 minute. The thermocycler is so fast that the DNA biochemistry is rate limiting. The PCRJet has been used to diagnose Bacillus anthracis and closely related B. cereus using 30-40 PCR cycles in 3.5 to 8.5 minutes. With engineering improvements and real-time detection optics, it should be possible to routinely amplify and detect special pathogens in < 8 minutes. In Phase II Research, a high performance PCRJet will be fabricated based on Computational Fluid Dynamic modeling. This machine will be capable of both high-speed DNA amplification and fluorescent dye-based detection, so that diagnostic B. anthracis and Variola major DNA fragments can be detected in ~5 minutes. Using a biochemical engineering approach to optimization of the PCR process, a very fast thermostable Jet Polymerase enzyme mixture (KOD Pol + dUTPase + PPase) will be formulated, so that elongation rates >500 nt/sec at 72xC can be achieved. The combination of the PCRJet thermocycler and improved enzymology for high-speed amplification of DNA will enable diagnosis of agents of bioterrorism on a time-scale of <2 minutes.

Principal Investigator:

Nisha V. Padhye
4024676499
NPADHYE@CVIENZYMES.COM

Business Contact:

R. Nelson
4024676499
MNELSON@CVIENZYMES.COM
Small Business Information at Submission:

MEGABASE RESEARCH PRODUCTS
MEGABASE RESEARCH PRODUCTS 2820 N 48TH ST, STE 110 LINCOLN, NE 68504

EIN/Tax ID: 470755195
DUNS: N/A
Number of Employees: N/A
Woman-Owned: No
Minority-Owned: No
HUBZone-Owned: No