Patent ID: 6270963
Filing Date: 2001-08-07
Classification: C12Q

Abstract:
A method for testing a plurality of patients for a disease-associated mutation in a gene of interest comprising the steps of:(a) performing an immunoassay on samples obtained from each of the plurality of patients by combining a portion of sample with an antibody which forms an immunological reaction product by binding to a protein gene product of the gene of interest and monitoring for formation of the immunological reaction product, said antibody being selected to provide fewer than 1% false results indicating the presence of a mutation when combined with the protein gene product of the gene of interest, whereby the formation of an immunological product is indicative of the presence or absence of a mutation in the gene of interest;(b) performing a probe-based assay on each patient sample for which the results of the immunoassay did not indicate the presence of a mutation, but not on those samples for which the immunoassay indicated the presence of the mutation by combining a second portion of each patient sample with a nucleic acid probe which specifically and selectively hybridizes with the gene of interest in its mutated or wild-type form, whereby the formation of a duplex nucleic acid hybrid containing the nucleic acid probe is indicative of the presence or absence, respectively, of the mutation in the gene of interest; and(c) determining the sequence of DNA in at least one selected region of the gene of interest on each patient sample for which the results of the probe-based assay did not indicate the presence of the mutation and comparing the sequence determined with known sequences of normal or mutant forms of the gene of interest,wherein the immunoassay and probe-based assay used are selected by a method comprising the steps of:identifying a plurality of immunoassays and probe-based assays for the disease-associated mutation;determining the level of false results for each assay by comparing results from assays on a statistically significant plurality of samples against results from assays on the same samples using a test protocol accepted as having essentially no false results;defining an outcome mapping matrix containing a plurality of diagnostic algorithms each comprising a combination of assays to be evaluated and assessing the significance of all possible combinations of negative and positive results for the algorithms in the matrix;determining overall sensitivity overall specificity, overall predictive value of a positive algorithm result and overall predictive value of a negative algorithm result for at least some of the algorithms in the outcome matrix;determining the estimated cost of testing for those combinations where the overall sensitivity overall specificity, overall predictive value of a positive algorithm result and overall predictive value of a negative algorithm result meet a predefined threshold; andselecting a diagnostic algorithm of assays based upon the low estimated cost of testing.