Patent ID: 7250252

Claim:
A method for detecting a target nucleic acid sequence suspected of having single or large deletions or insertions in a test sample comprising the steps of: a) contacting the test sample with amplification reagents comprising a polymerase, a primer pair, and a probe to form a reaction mixture; b) performing the following cycle comprising the steps of: (i) maintaining the reaction mixture for a time and at temperature above 90° C., sufficient to dissociate double stranded nucleic acid sequences, (ii) maintaining the reaction mixture for a time and at a temperature from 45° C. to 65° C. to allow the primers and probe to hybridize to the nucleic acid and thereby form primer hybrids and probe hybrids, (iii) maintaining the reaction mixture for a time and at a temperature at least 1° C. above the temperature in (ii), sufficient to dissociate the probe hybrids, if the probe is not completely complementary to the nucleic acid, and (iv) raising the temperature of the reaction mixture to a temperature sufficient to activate the polymerase; c) repeatedly performing the cycle of step b) to form an amplification product; and d) detecting the amplification product as an indication of the presence of the nucleic acid sequence in the test sample.