Patent ID: 9284607
Filing Date: 2016-03-15
CPC Classification: C12Q,C40B

Claim Text:
1. A method for assaying a sample for one or more target nucleic acids, said method comprising: (a) contacting a sample comprising one or more target nucleic acids with a polymerase chain reaction mixture comprising: (b) performing a polymerase chain reaction on the sample/reaction mixture under amplification conditions, wherein, when a target nucleic acid is present, THO which are substantially complementary to part of that target nucleic acid are hybridised with the target nucleic acid, therefore being consumed, wherein the consumption of probes causes changes of detectable signal in the labels, and the consumed probes are no longer able to form a double stranded portion if the original probe has a double-stranded portion; and (c) measuring, at least once, the melting profile of the unconsumed probes in the reaction mixture by detecting the signal(s) from the labels in those probes as a function of temperature, wherein the presence or absence of melting characteristics of any probes in the melting profile analysis is an indication of unconsumption or consumption of that probe, which further provides an indication of whether or not at least one target nucleic acid is present in said sample, wherein THO is complementary to a target sequence, and is labeled with a fluorophore and a quencher, wherein PCO, which is partially complementary to THO, contains modified the 3′end to prevent its extension, wherein, if the 3′ end of PCO is attached with a label which is not quencher, fluorescence emission is increased by hybridisation of THO and PCO, this type of probe is termed plus probe (+THO:PCO), wherein, if the 3′ end of PCO is attached with a quencher, fluorescence emission is decreased by hybridisation of THO and PCO, this type of probe is termed minus probe (−THO:PCO).