| 摘要 |
<p>PURPOSE: A nucleic acid isothermal amplification method of a non-temperature controlling method using a heat resistant single strand binding protein and a heat resistant helicase II, and a nucleic acid detection method using the same are provided to detect amplification of a target nucleic acid on a real-time basis through amplification of a probe signal in a high temperature isothermal condition by using a SSB protein. CONSTITUTION: An isothermal amplification method of a high temperature nucleic acid comprises a step of preparing reaction solution by mixing a primer pair, a heat resistant helicase II, a heat resistant SSB protein, and polymerase to a sample which includes a target nucleic acid; and a step of amplifying the target nucleic acid in a isothermal state of 50-75°C. The heat resistant helicase II has an amino acid sequence of sequence number 11. A heat resistant SSB protein has amino acid sequence of sequence number 13. The step of amplifying amplifies at 50-70°C. The isothermal detecting method of the high temperature nucleic acid comprises a step of preparing reaction solution by mixing a primer pair for target amplification, a fluorescent label-probe, helicase II, heat resistant SSB protein, and polymerase to a sample which includes a target nucleic acid; and a step of amplifying the target nucleic acid in an isothermal state of 50-75°C. A fluorescent label-probe has a quencher or a fluorescent protein or a quencher in the 5' end part thereof, and a fluorescence protein or a quencher in the 3' end part thereof.</p> |
