Abstract: This specification relates to non-radioactive methods, compositions and kits for non-radioactive real-time PCR using FRET dual-labeled primers and do not require the use of a probe. The non-radioactive methods may also be used for end-point PCR in which the signal is measured only at the endpoint of the PCR cycling. The dual-labeled primer maintains the molecular tether between fluorophore and quencher. Kits are also provided for the quantification or detection of one or more target nucleic acid molecules in a sample during nucleic acid synthesis, and include a dual-labeled oligonucleotide.

Abstract: This specification relates to non-radioactive methods, compositions and kits for non-radioactive real-time PCR using FRET dual-labeled primers and do not require the use of a probe. The non-radioactive methods may also be used for end-point PCR in which the signal is measured only at the endpoint of the PCR cycling. The dual-labeled primer maintains the molecular tether between fluorophore and quencher. Kits are also provided for the quantification or detection of one or more target nucleic acid molecules in a sample during nucleic acid synthesis, and include a dual-labeled oligonucleotide.

Abstract: The present disclosure, in some embodiments, relates to compositions and methods for detection of multiple target nucleic acid sequences by a single nucleic acid amplification based assay (e.g. PCR). Compositions comprising two or more primers are described for multiple target nucleic acid detection. Methods for differential detection of microorganisms (including strains/serovars/subtypes thereof) and cell types comprise a single step method to detect a signature of target nucleic acid sequences comprising: two, three or more target nucleic acids that are uniquely present in a microorganism/strain/serovar/subtype and/or cell and absent in other closely related organisms/cells. Embodiments relate to methods of diagnosis of diseases or conditions that can be detected by detecting the presence of two or more nucleic acid target markers.

Abstract: This specification generally relates to non-radioactive methods of non-radioactive real-time PCR using FRET dual-labeled primers.