DIAGNOSTIC AND AMPLIFICATION METHODS OVERCOMING A PRIMER-TARGET MISMATCH AT THE PRIMER 3' END

摘要

DIAGNOSTIC AND AMPLIFICATION METHODS OVERCOMING A PRIMER-TARGET MISMATCH AT THE PRIMER 3' END Methods for amplifying and detecting a predetermined target nucleic acid in a biological specimen are accomplished even where there is a mismatch in a single position between a primer and the target nucleic acid. The mismatch is located at or near the 3' end of the primer. Such a mismatch is overcome using a primer having a nucleotide with a thymine base at the position of the mismatch. The use of such primers is most likely to prime the target and form primer extension products. This method is particularly useful for detection of a nucleic acid sequence which is not fully known, or where there is considerable heterogeneity in DNA target from patient samples.