Molecular Diagnostic Assays for Genetic Disease

DESCRIPTION (Applicant's Abstract): The objective of Phase 1 is to develop a
sensitive test and kit for detection of the Factor V Leiden mutation in human
blood using a new nucleic acid ligation-based technology invented at Lumigen.
The ligation technology is based on the discovery that a multitude of short
oligonucleotides can be contiguously ligated to a template-bound anchor
sequence performed under conditions in which the short oligonucleotides do not
stably hybridize to the template. The absence of stable hybridization of the
short oligonucleotides during the ligation confers extremely high replication
fidelity. Incorporation of detectable labels on some or all of the short
oligonucleotides permits the design of molecular assays with excellent
specificity and reliability of detection. We will use a small hapten label to
be detected with an enzyme-antibody conjugate and a chemiluminescent enzyme
substrate.
PROPOSED COMMERCIAL APPLICATION:
The proposed test methods and kits should provide the basis for developing a set of
detection methods and products for analysis of many different SNPs. The tests involve a
new amplification process which should be more specific than existing amplification
technology while achieving comparable speed and sensitivity. The tests should be capable
of automation on a high throughput platform including chip formats.
The unique properties of this methodology have resulted in the creation of a
new nucleic acid amplification technology which is more specific than PCR and
yields cleaner products. We will use this amplification technique to enhance
the sensitivity of the method to detect the single-nucleotide polymorphism
(SNP) associated with this mutation. The feasibility of developing a single
test for discrimination of all three possible genotypes of this mutation will
also be explored. This test will utilize one of two patented methods developed
at Lumigen for detection of two enzyme labeled analytes using proprietary
chemiluminescent substrates.