Isothermal amplification components and processes

A method for determining the presence, absence or amount of a target sequence in sample nucleic acid, comprising:

• a) amplifying a target sequence in the sample nucleic acid, wherein:
• the target sequence comprises a first strand and a second strand,
• the first strand and second strand are complementary to each other, and
• the amplifying comprises contacting non-denatured sample nucleic acid under cleavage agent-free, helicase-free and recombinase-free isothermal amplification conditions with:
• i) a first oligonucleotide and a second oligonucleotide, wherein:
• the first oligonucleotide consists of a first polynucleotide continuously complementary to a sequence in the first strand, and
• the second oligonucleotide consists of a second polynucleotide continuously complementary to a sequence in the second strand;
• and
• ii) at least one component providing a hyperthermophile polymerase activity, thereby generating a nucleic acid amplification product, wherein:
• the nucleic acid amplification product consists of 1) a first nucleotide sequence that is continuously complementary to or substantially identical to the first polynucleotide of the first oligonucleotide, 2) a second nucleotide sequence that is continuously complementary to or substantially identical to the second polynucleotide of the second oligonucleotide, and 3) a spacer sequence comprising 1 to 10 bases;
• and
• the spacer sequence is flanked by the first nucleotide sequence and the second nucleotide sequence;
• and
• b) detecting the nucleic acid amplification product, wherein detecting the nucleic acid amplification product comprises use of a real-time detection method and is performed in 10 minutes or less from the time the sample nucleic acid is contacted with (a)(i) and (a)(ii), whereby the presence, absence or amount of a target sequence in sample nucleic acid is determined.