Process for lysing mycobacteria
AssigneeBecton, Dickinson and Company
InventorsJames A. Down, Melinda S. Fraiser, G. Terrance Walker
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Abstract
A process for lysing Mycobacteria to liberate nucleic acids such as DNA and RNA comprises heating the Mycobacteria in an aqueous solution to lyse the Mycobacteria and release the nucleic acids, with the aqueous solution containing a chelating agent such as EDTA or EGTA in an amount effective to inhibit degradation of the released nucleic acids. Examples of Mycobacteria which can be lysed include Mycobacterium fortuitum and Mycobacterium tuberculosis. After lysis, the nucleic acid is preferably then amplified and detected. Kits useful for carrying out the present invention are also disclosed.
Claims16
A process for amplifying a Mycobacteria nucleic acid segment in a sample suspected of containing Mycobacteria, comprising:
- combining said sample with a lysis solution consisting essentially of less than about 10 mM EGTA in water or a nucleic acid-compatible buffer;
- heating said lysis solution for a time and to a temperature effective to lyse Mycobacteria present therein;
- and
- amplifying the Mycobacteria nucleic acid segment in a DNA amplification reaction in said lysis solution.
A kit useful for amplifying Mycobacteria nucleic acid in a, comprising:
- (a) a lysis solution consisting essentially of less than 10 mM EGTA in water or a nucleic acid-compatible buffer;
- and
- (b) a nucleic acid amplification reagent.
Dependencies are inferred from the claim text.