Methods and apparatus for screening and detecting multiple genetic mutations

摘要

An assay system and methods are described where patient samples containing genomic DNA are analyzed for the presence of known genetic polymorphisms using a universal reporter strategy. In a preferred embodiment, the amplified DNA is localized at test sites in an array of sites on a microchip followed by a series of hybridization reactions that screen for the presence of a single mutation from among a number of mutations, and allow the identification of specific mutations. In addition to universal reporters, the assay may use blockers and discriminators for screening and identification of known polymorphisms.

主权项

1. A kit for use in a process of detecting members of a set of polymorphisms that occur at identified loci in samples of a patient nucleic acid, said kit comprising:
a group of unlabeled blockers capable of hybridizing under stringent conditions with markers in at least one locus contained in the patient sample nucleic acid thereby blocking said markers from further hybridizing to a detectable discriminator, wherein a specific nucleotide sequence of at least one unlabeled blocker of said group of unlabeled blockers is fully complementary to a nucleotide sequence comprising a mutation or a polymorphism in said at least one locus, and wherein the at least one unlabeled blocker is capable of hybridizing under said stringent conditions to the nucleotide sequence comprising the mutation or the polymorphism; more than one detectable discriminator, at least one of said more than one detectable discriminator capable of hybridizing under said stringent conditions with the nucleotide sequence comprising said mutation or polymorphism, said at least one unlabeled blocker being capable of blocking said at least one detectable discriminator from hybridizing with the nucleotide sequence comprising said mutation or polymorphism under said stringent conditions; and at least one universal reporter capable of binding with said more than one detectable discriminator, wherein said at least one of said more than one detectable discriminator is denatured from the nucleotide sequence comprising said mutation or said polymorphism in a hybridization product formed by said at least one detectable discriminator and the nucleotide sequence comprising said mutation or said polymorphism under denaturation condition while said at least one unlabeled blocker has a sufficient length so that said at least one unlabeled blocker remains to hybridize with the nucleotide sequence comprising said mutation or said polymorphism in a hybridization product formed by said at least one unlabeled blocker and the nucleotide sequence having said mutation or said polymorphism under said denaturation condition.