A -- DNA MUTATION TESTS

Notice Date

July 12, 2001

Contracting Office

NCI Frederick Cancer Research and Development Center (NCI-FCRDC), P.O. Box B, Frederick, Maryland 21702-1201

ZIP Code

21702-1201

Response Due

August 13, 2001

Point of Contact

Patricia L. Getzandanner, Sr. Contract Specialist, 301-846-1721, Contracting Officer -- Dennis J. Dougherty, 301-846-1087

E-Mail Address

Patricia L. Getzandanner, Sr. Contract Specialist (pgetzandanner@mail.ncifcrf.gov)

Description

Introduction: R&D -- Potential Sources Sought. The National Cancer Institute (NCI) is initiating a new program to study inherited bone marrow failure syndromes (BMFS) in children. Fanconi's Anemia (FA) is the prototype of these diseases, but the study will not be limited to this one syndrome. There are seven genes known to play an etiologic role in FA, with most of the mutations occurring in the Fanconi "A" gene. There is also a founder mutation in the FA "C" gene that accounts for many of the cases that arise in a specific subset of individuals. The genes are large and there are multiple mutations within each gene. A contractor is sought to develop appropriate DNA mutation tests to provide genotype data for genotype/phenotype/cancer susceptibility risk factor analysis. 1. Laboratory components: 1) Testing for Fanconi Anemia Group A (FANCA): a) Design, develop, acquire and test the estimated 40-50 primers required for DNA amplification of the FANCA gene b) Develop the specific DNA sequence analysis and DHPLC methods required for sequencing of this gene c) Extract mRNA from submitted samples d) Prepare and amplify the entire cDNA by RT-PCR e) Perform bi-directional sequence analysis of the entire cDNA, to identify missense and frame-shift mutations that account for approximately half of the FANCA mutations f) Agarose gel analysis of cDNA to identify samples that show size differences due to splice-site mutations, followed by direct gDNA sequence analysis of intron/exon boundaries represented by an abnormally sized cDNA fragment g) Quantitative multiplex PCR/DHPLC assay to detect large multi-exon deletions in genomic DNA 2) Testing for Fanconi Anemia Group C (FANCC, IVS4AtoT), Ashkenazi Jewish mutation, using specific PCR primers 3) Testing for Fanconi Anemia Group C (FANCC) other mutations, and for Fanconi Anemia other Groups (FANCD-G): a) Methods used for FANCA or for specific mutations, as appropriate on a gene by gene basis. 4) Testing for Diamond-Blackfan Anemia (DBA) RPS19 gene: a) Sequence the 5 exons of RPS19 5) Testing for Dyskeratosis Congenita (DC) Gene DKC1: a) Sequence the entire DKC1 gene cDNA b) Sequence genomic DNA as needed 6) Testing for Severe Congenital Neutropenia (SCN) Gene, neutrophil elastase 2 (ELA2): a) Sequence the 5 exons of ELA2 7) Testing for Amegakaryocytic Thrombocytopenia (Amega) Gene, MPL: a) Sequence the entire gene cDNA 8) Testing for Pearson's Syndrome Gene Deletion in Mitochondrial DNA: a) Identify mutations by gel electrophoresis of mitochondrial DNA 9) Testing for other mutations in bone marrow failure syndromes, which have yet to be cloned: a) Using the most appropriate techniques, similar to those outlined above. There will be approximately 35 samples to be tested for the FANCA during the first year, and five or less for each of the other mutations. Written reports for each analysis will be required. Laboratories interested in participating must be CLIA certified. Organizations interested in participating are requested to contact Patricia Getzandanner, Sr. Contract Specialist, #301-846-1721, or e-mail at pgetzandanner@mail.ncifcrf.gov

Record

Loren Data Corp. 20010716/ASOL007.HTM (W-193 SN50R6N5)