A System for Rapid PCR, Mutation Scanning and Genotyping

Award Information
Agency: Department of Health and Human Services
Branch: N/A
Contract: 4R42GM073396-02
Agency Tracking Number: GM073396
Amount: $750,000.00
Phase: Phase II
Program: STTR
Awards Year: 2005
Solicitation Year: N/A
Solicitation Topic Code: N/A
Solicitation Number: N/A
Small Business Information
390 Wakara Way, Salt Lake City, UT, 84108
DUNS: N/A
HUBZone Owned: N
Woman Owned: N
Socially and Economically Disadvantaged: N
Principal Investigator
 CARL WITTWER
 (801) 581-4737
 CARL.WITTWER@PATH.UTAH.EDU
Business Contact
 MONTY BOTOSAN
Phone: (801) 582-5138
Email: MONTY@IDAHOTEC.COM
Research Institution
N/A
Abstract
DESCRIPTION (provided by applicant): Our objective is to provide a closed-tube system for rapid PCR, mutation scanning and genotyping that does not require fluorescently-labeled probes. For many genetic diseases, it is difficult and expensive to screen for all possible mutations that may cause the disease. We propose a relatively simple solution for rapid amplification, scanning and genotyping in a single homogeneous system. Certain DNA dyes are compatible with PCR and can detect mismatches between two copies of DNA by simple melting analysis after amplification. The dye is added before PCR, and heterozygotes are easily identified after a 1-2 min high-resolution melting curve after PCR. Specific genotype confirmation is provided by an unlabeled oligonucleotide probe. The specific aims for Phase I of this Fast Track proposal are: 1. Develop a homogeneous, closed-tube gentotyping method that uses unlabeled oligonucleotides. 2. Prototype a rapid PCR system that integrates high-resolution melting analysis for scanning and genotyping. We will integrate the high-resolution melting capability of our HR-1 instrument into our rapid PCR machine (R.A.P.I.D./LightCycler). After PCR, each tube will be automatically analyzed by high-resolution melting. Simultaneous scanning and genotyping will be demonstrated using special DNA dyes. Advantages of our system includes speed (PCR in 15 min, analysis in 1-2 min/sample), homogeneous design (no need for sample transfer or reagent additions), closed-tube analysis (no amplicon contamination risk), both scanning and genotyping capability, and nondestructive analysis (immediately available for sequencing in the rare case when it is necessary).

* Information listed above is at the time of submission. *

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