Nucleic acid amplification and testing
AssigneeCambridge Enterprise Limited
InventorsHelen Hwai-an Lee, Magda Anastassova Dineva, Fiona Frances Sarah Fletcher-Brown
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Abstract
Methods for amplifying a target nucleic acid by self-sustained amplification methods are described. The methods are designed, in particular, to be carried out without use of specialised lab facilities or instruments. Compositions, lyophilised formulations, and kits for carrying out the methods are also described.
Claims46
A method of nucleic acid amplification, which comprises amplifying a target nucleic acid by a self-sustained transcription-based amplification reaction which is carried out at a temperature above 42° C. and less than 50° C.
A method of testing for the presence of a target nucleic acid in a sample solution suspected of containing the target nucleic acid, the method comprising incubating the sample solution under conditions for amplification of target nucleic acid by a self-sustained transcription-based amplification reaction at a temperature above 42° C. and less than 50° C.
A method according to any preceding claim, wherein at least one non-thermostable enzyme is used.
A method according to any preceding claim, which is carried out without pre-incubating the target nucleic acid or sample solution at a temperature of 50° C. or above.
A method according to any preceding claim, wherein the amplification reaction is carried out, or the sample solution is incubated under conditions for amplification of target nucleic acid, in a processing chamber that is sealed from the external environment.
A method according to any preceding claim, which further comprises detecting for product resulting from amplification of the target nucleic acid.
A method according to any preceding claim, wherein reagents required for amplification of the target nucleic acid and/or detection of product resulting from amplification of the target nucleic acid are provided in lyophilised form.
A method according to any preceding claim, wherein the amplification reaction is carried out in the presence of a polysaccharide and a low molecular weight saccharide.
A lyophilised formulation which comprises a polysaccharide, a low molecular weight saccharide, and a labile reagent which it is desired to preserve in a stable condition, wherein the formulation does not include a buffering agent, or the buffering agent is present at a concentration of less than 1 mM.
A lyophilised formulation, which comprises a polysaccharide, a low molecular weight saccharide, and an antibody (or a fragment or derivative thereof that retains antigen binding activity), in the absence of an antigen bound by the antibody.
A primer or probe that comprises or consists of a sequence recited in any of SEQ ID NOs:
- 1 to 9.
A set of primers that comprises a primer with sequence recited in SEQ ID NO:
- 1 and a primer with sequence recited in SEQ ID NO:
- 2.
A set of primers that comprises a primer with sequence recited in SEQ ID NO:
- 6 and a primer with sequence recited in SEQ ID NO:
- 7.
A set of primers and probes that comprises:
- a primer with sequence recited in SEQ ID NO:
- 1;
- a primer with sequence recited in SEQ ID NO:
- 2;
- a probe with sequence recited in SEQ ID NO:
- 3;
- a probe with sequence recited in SEQ ID NO:
- 4;
- and
- a probe with sequence recited in SEQ ID NO:
- 5.
A set of primers and probes that comprises:
- a primer with sequence recited in SEQ ID NO:
- 6;
- a primer with sequence recited in SEQ ID NO:
- 7;
- a probe with sequence recited in SEQ ID NO:
- 8;
- and
- a probe with sequence recited in SEQ ID NO:
- 9.
Dependencies are inferred from the claim text.