METHOD OF AMPLIFYING AND DETECTING TARGET NUCLEIC ACID SEQUENCE BY USING THERMOSTABLE ENZYMES.

摘要

A method of amplifying a specified nucleic acid sequence, which permits an amplification reduced in nonspecific hybridization and increased in specificity. The invention also provides a reagent which does not suffer any activity drop during supply and preservation thereof and is highly stable. Thermostable enzymes are used as the RNA-dependent DNA polymerase, DNA-dependent DNA polymerase, DNA-dependent RNA polymerase and ribonuclease H, all of which are necessary for the replicative RNA-base amplification system. It is desirable to use combinedly a thermostable DNA-dependent RNA polymerase and a thermostable enzyme originating in Thermus thermophilus which comprises all of the RNA-dependent DNA polymerase, DNA-dependent DNA polymerase and ribonuclease H. The use of the thermostable enzymes according to the above method serves to prevent the enzymes from being deactivated and enables the amplification without the necessity for sequentially adding the enzymes. <IMAGE>