DETECTING SINGLE NUCLEOTIDE POLYMORPHISM USING OVERLAPPING HYDROLYSIS PROBES

摘要

Methods for the rapid detection of the presence or absence of a SNP in a target nucleic acid in a sample are described. The methods can include performing an amplifying step, a hybridizing step utilizing a double stranded probe with two overlapping SNP specific hydrolysis probe sequences where one of the probe sequences can include a hairpin structure toward the 3′ end, and a detecting step. Furthermore, the double stranded SNP specific hydrolysis probes along with kits are provided that are designed for the detection of a SNP in a target nucleic acid.

主权项

1. A method for detecting a single nucleotide polymorphism (SNP) in a target nucleic acid in a sample, the method comprising:
performing an amplifying step comprising contacting the sample with a first primer comprising a first nucleic acid and a second primer comprising a second nucleic acid sequence to produce an amplification product comprising a sense strand and an anti-sense strand if any target nucleic acid is present in the sample; performing a hybridizing step comprising providing the amplification product with a double stranded probe comprising:
a first SNP specific hydrolysis probe comprising a third nucleic acid sequence complementary to a first SNP containing region of the sense strand, the first SNP specific hydrolysis probe comprising a first interactive label and a second interactive label, a first 5′ end and a first 3′ end; anda second SNP specific hydrolysis probe comprising a fourth nucleic acid sequence comprising a plurality of nucleotides complementary to a SNP containing region of the anti-sense strand, the second SNP specific hydrolysis probe comprising a third interactive label and a fourth interactive label, a second 5′ end and a second 3′ end; and detecting the presence or absence of the amplification product, wherein the presence of the amplification products is indicative of the presence of the SNP in the target nucleic acid target, and wherein the absence of the amplification products is indicative of the absence of the SNP in the target nucleic acid target.