METHOD FOR CONCENTRATING VARIANT NUCLEIC ACID AND NUCLEIC ACID CONCENTRATION TEST KIT FOR EFFECTING THE METHOD

摘要

A method for selectively concentrating a mutant nucleic acid constituting a particular segment of a target nucleic acid is provided, and this method is characterized in that: a cycle comprising the steps of (1) to (3):(1) the step of preparing a nucleic acid sample by amplifying said particular segment of said target nucleic acid,(2) the step of adding to said nucleic acid sample an equimolar amount or more of a labeled nucleic acid standard prepared by incorporating a label capable of binding to a solid support into a nucleic acid having a nucleotide sequence complementary to the normal nucleic acid sequence of said particular segment of said target nucleic acid to promote competitive hybridization, and(3) the step of separating and removing the hybridizate having said label capable of binding to the solid support and the residual labeled nucleic acid standard in the reaction solution after the competitive hybridization by trapping them on the solid support; ora cycle comprising the steps of (1') to (4'):(1') the step of preparing a labeled nucleic acid sample by amplifying said particular segment of said target nucleic acid, and simultaneously, incorporating two types of labels respectively capable of binding to solid supports into the amplification product,(2') the step of adding to said labeled nucleic acid sample an equimolar amount or more of a nucleic acid standard having a nucleotide sequence complementary to the normal nucleic acid sequence of said particular segment of said target nucleic acid to promote competitive hybridization,(3') the step of collecting the hybridizates having one of said two types of labels from the reaction solution after the competitive hybridization by trapping such hybridizates on the first solid support which selectively binds to said one of said two labels, and(4') the step of collecting the hybridizate having both of said two types of labels or a single strand nucleic acid derived from said hybridizate by contacting the hybridizates collected in the step (3') with the second solid support which selectively binds to the other one of the two labels is effected one or more times; or said cycle is effected once and said steps (2) and (3) or said steps (2') to (4') are repeated one or more times.