A method for simultaneous multiplex real-time PCR amplification, detection and quantification of one or more target nucleic acids in a single sample container comprising the steps of: (a) contacting a sample, suspected of containing one or more target nucleic acids, with at least one set of oligonucleotide probes, each oligonucleotide probe within the set labeled with the same one or more reporter moieties, wherein each of said labeled oligonucleotide probes i. is sufficiently complementary to at least a subsequence of at least one target nucleic acid; ii. is capable of binding to the corresponding target nucleic acid with a melting temperature distinct from the melting temperatures of the other labeled oligonucleotide probes within the same set; (b) amplifying the target nucleic acids in the sample in a cycle of an amplification reaction that includes a temperature change interval, wherein said labeled oligonucleotide probes dissociate from the hybrids with the corresponding target nucleic acids; (c) detecting light emission from said reporter moieties over at least a portion of said temperature change interval; and (d) plotting the first derivative of said light emission detected in step (c) over at least said portion of the temperature change interval; (e) determining the maximum value of said derivative plotted in step (d); (f) repeating steps (b) through (e) multiple times; and (g) plotting the maximum values of said derivative determined in step (e) against the cycle number or the number of repetitions of steps (b) through (e), and determining the cycle number or the number of repetitions at which a predetermined threshold value of the value determined in step (e) is reached, thus quantifying the relative amount of said target nucleic acid, wherein said simultaneous multiplex real-time PCR amplification, detection and quantification reaction comprises the detection of at least two different target sequences either comprising amplification using separate pairs of amplification primers for each target sequence and detection using a separate oligonucleotide probe for each target sequence or amplification using a pair of amplification primers and detection using more than one oligonucleotide probe for each amplified sequence. (a) contacting a sample, suspected of containing one or more target nucleic acids, with at least one set of oligonucleotide probes, each oligonucleotide probe within the set labeled with the same one or more reporter moieties, wherein each of said labeled oligonucleotide probes i. is sufficiently complementary to at least a subsequence of at least one target nucleic acid; ii. is capable of binding to the corresponding target nucleic acid with a melting temperature distinct from the melting temperatures of the other labeled oligonucleotide probes within the same set; i. is sufficiently complementary to at least a subsequence of at least one target nucleic acid; ii. is capable of binding to the corresponding target nucleic acid with a melting temperature distinct from the melting temperatures of the other labeled oligonucleotide probes within the same set; (b) amplifying the target nucleic acids in the sample in a cycle of an amplification reaction that includes a temperature change interval, wherein said labeled oligonucleotide probes dissociate from the hybrids with the corresponding target nucleic acids; (c) detecting light emission from said reporter moieties over at least a portion of said temperature change interval; and (d) plotting the first derivative of said light emission detected in step (c) over at least said portion of the temperature change interval; (e) determining the maximum value of said derivative plotted in step (d); (f) repeating steps (b) through (e) multiple times; and (g) plotting the maximum values of said derivative determined in step (e) against the cycle number or the number of repetitions of steps (b) through (e), and determining the cycle number or the number of repetitions at which a predetermined threshold value of the value determined in step (e) is reached, thus quantifying the relative amount of said target nucleic acid,