PROCEDURE FOR DETECTING TARGET NUCLEIC ACID AFTER AMPLIFICATION

摘要

PROBLEM TO BE SOLVED: To detect a target nucleic acid for diagnosis of gene disturbance and infectious diseases by amplifying a specimen in which the presence of the target nucleic acid is doubtful with a primer complementary to the target nucleic acid and a thermally stable amplification enzyme and by measuring the nucleic acid after modification and deactivation of the enzyme. SOLUTION: A specimen in which the presence of a target nucleic acid is doubtful is brought into contact with a primer comprising at least two kinds of oligonucleotides which are substantially complementary to a part of the target nucleic acid and a thermally stabilized DNA polymerase as a thermally stable amplification enzyme under such conditions as this target nucleic acid may be amplified. Then, the amplified target nucleic acid is modified to form a single-stranded nucleic acid and the reaction mixture containing the specimen is incubated at 95-120 deg.C for 1 second to 30 minutes to deactivate the thermally stable amplification enzyme and whether this target nucleic acid is present or not is detected, whereby a target nucleic acid for a variety of medical and academic researches, forensic medical examinations and diagnosis of gene disturbance and infectious diseases can be detected through amplification.