Processes for detection of nucleic acids

A process for detecting qualitatively or quantitatively the presence of a single-stranded or double-stranded nucleic acid of interest in a sample, said process comprising the steps of:

• (a) providing
• (i) a sample suspected of containing said nucleic acid of interest;
• (ii) a first nucleic acid primer that comprises a nucleic acid sequence complementary to at least a portion of one strand of said nucleic acid of interest;
• (iii) a second nucleic acid primer that comprises a nucleic acid sequence identical to at least a portion of said one strand;
• (iv) labeled nucleotide or labeled nucleotides comprising a first energy transfer element;
• and
• (v) reagents for carrying out nucleic acid strand extension;
• wherein
• the first nucleic acid primer, the second nucleic acid primer, or both the first nucleic acid primer and the second nucleic acid primer comprise a second energy transfer element, and
• said first energy transfer element is an energy transfer donor and said second energy transfer element is an energy transfer acceptor, or said first energy transfer element is an energy transfer acceptor and said second energy transfer element is an energy transfer donor;
• (b) forming a reaction mixture comprising (i), (ii), (iii), (iv) and (v) above, wherein said reaction mixture lacks the presence of any nucleotides that are strand terminators;
• (c) contacting under hybridization conditions said first nucleic acid primer with one strand of said nucleic acid of interest and contacting under hybridization conditions said second nucleic acid primer with the complementary strand of said nucleic acid of interest if present;
• (d) extending said first nucleic acid primer and said second nucleic acid primer to form a first primer-extended nucleic acid sequence and a second primer-extended nucleic acid sequence if said complementary strand is present, thereby incorporating more than one labeled nucleotides into each of the extended primers;
• (e) separating said first primer-extended nucleic acid sequence from said nucleic acid of interest and separating said second primer-extended nucleic acid sequence from said complementary strand of said nucleic acid of interest if present;
• (f) contacting under hybridization conditions said first nucleic acid primer with said nucleic acid of interest or said second primer-extended nucleic acid sequence from step (e), and contacting under hybridization conditions said second nucleic acid primer with said first primer-extended nucleic acid sequence from step (e);
• and
• (g) detecting the presence or quantity of said nucleic acid of interest by detecting energy transfer between a second energy transfer element in said first nucleic acid primer, said second nucleic acid primer, or both, and a first energy element in an incorporated nucleotide.

A process for detecting qualitatively or quantitatively the presence of a single-stranded or double-stranded nucleic acid of interest in a sample, said process comprising the steps of:

• (a) providing
• (i) a sample suspected of containing said nucleic acid of interest;
• (ii) a first nucleic acid primer that comprises a nucleic acid sequence complementary to at least a portion of one strand of said nucleic acid of interest;
• (iii) a second nucleic acid primer that comprises a nucleic acid sequence identical to at least a portion of said one strand;
• (iv) one or more first nucleotides, said first nucleotides comprising a first energy transfer element, and one or more second nucleotides, said second nucleotides comprising a second energy transfer element, wherein said first energy transfer element is an energy transfer donor and said second energy transfer element is an energy transfer acceptor, or said first energy transfer element is an energy transfer acceptor and said second energy transfer element is an energy transfer donor;
• and
• (v) reagents for carrying out nucleic acid strand extension;
• (b) forming a reaction mixture comprising (i), (ii), (iii), (iv) and (v) above, wherein said reaction mixture lacks the presence of any nucleotides that are strand terminators;
• (c) contacting under hybridization conditions said first nucleic acid primer with one strand of said nucleic acid of interest and contacting under hybridization conditions said second nucleic acid primer with the complementary strand of said nucleic acid of interest if present;
• (d) extending said first nucleic acid primer and said second nucleic acid primer in a PCR reaction to form a first primer-extended nucleic acid sequence and a second primer-extended nucleic acid sequence if said complementary strand is present, thereby incorporating said first and second nucleotides;
• and
• (e) detecting the presence or quantity of said nucleic acid of interest by detecting energy transfer between said first energy transfer element and said second energy transfer element during said PCR reaction.