METHOD AND COMPOSITIONS FOR PRIMER SPECIFIC SOLID-PHASE DETECTION OF PCR PRODUCTS

摘要

The invention provides a method of detecting the presence of a nucleic acid in a sample comprising the following steps: a) amplifying a nucleic acid from the sample using a first primer having a known sequence which can be selectively digested when in double stranded form and a second primer having a marker molecule such that a double stranded nucleic acid amplification product is formed having the known nucleotide sequence on one strand and the marker molecule on the other strand; b) digesting the known nucleotide sequence from one strand of the double stranded amplification product to expose a single stranded sequence complementary to the known nucleotide sequence on the strand having the marker molecule; c) hybridizing the single stranded sequence to a complementary nucleic acid bound to a solid support to capture the amplification product; d) removing uncaptured amplification product; and e) detecting the presence of the marker molecule captured on the solid support, the presence of marker molecule indicating the presence of the nucleic acid in the sample. Also provided is a kit comprising a pair of polymerase chain reaction primers, the first primer having a poly U tail on the 5' end and the second primer having a marker molecule linked to the 5' end. The kit can further include uracil DNA glycosylase. The kit can have biotin as the marker molecule and include streptavidin-horseradish peroxidase and a substrate for the horseradish peroxidase to detect the marker.