Sample pre-treatment for isothermal amplification
AssigneeNederlandse Organisatie voor toegepast-natuurwetenschappelijk onderzoek TNO
InventorsBart Jan Frederik KEIJSER, Michel OSSENDRIJVER
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Abstract
The invention is directed to a method for amplifying and/or detecting nucleic acid in a sample having a specific pre-treatment step. In particular, the invention provides a pre-treatment for the LAMP method, which can further enhance the sensitivity of LAMP tests. The pre-treatment includes a heat-treatment wherein the sample is heated in the presence of tris(2-carboxyethyl)phosphine (TCEP) and proteinase K.
Claims18
A method for amplifying nucleic acid, comprising the steps of
- (i) providing a sample comprising nucleic acid;
- and
- (ii) subjecting the sample to a heat-treatment in the presence of tris(2-carboxyethyl)phosphine (TCEP) and proteinase K;
- and
- (iii) an amplification step, wherein the heat-treated sample is subjected to loop-mediated isothermal amplification.
A method for detecting viral RNA from a sample, comprising the steps of
- (i) providing a sample comprising nucleic acid;
- and
- (ii) subjecting the sample to a heat-treatment in the presence of tris(2-carboxyethyl)phosphine (TCEP) and proteinase K;
- and
- (iii) an amplification step, wherein the heat-treated sample is subjected to loop-mediated isothermal amplification;
- and
- (iv) detecting the amplified viral RNA.
A pre-treatment for inactivating a sample comprising nucleic acid, comprising subjecting the sample to a heat-treatment in the presence of tris(2-carboxyethyl)phosphine (TCEP) and proteinase K.
A kit for treating a sample comprising nucleic acid to be amplified, comprising
- a buffer comprising TCEP and optionally EDTA;
- and
- a proteinase K solution;
- and
- a DNA polymerase and/or one or more LAMP primers
Dependencies are inferred from the claim text.