초록 ▼

The invention relates to methods and oligonucleotide probe compositions useful for determining antibiotic resistance in Mycobacteria. Included are methods for freeing intact precursor ribosomal RNA from mycobacterial cells and for assaying the levels of pre-rRNA in the cells. Also claimed are method

The invention relates to methods and oligonucleotide probe compositions useful for determining antibiotic resistance in Mycobacteria. Included are methods for freeing intact precursor ribosomal RNA from mycobacterial cells and for assaying the levels of pre-rRNA in the cells. Also claimed are methods useful in discovering new anti-mycobacterial therapeutic agents.

대표청구항 ▼

[ What is claimed is:] [1.] A method for detecting pre-ribosomal RNA (pre-rRNA) in cells of a mycobacterial sample, the method comprising the steps of:(a) treating the cells to release pre-rRNA, wherein said treatment comprises the steps of:(i) pretreating the cells by enzymatic degradation using bo

[ What is claimed is:] [1.] A method for detecting pre-ribosomal RNA (pre-rRNA) in cells of a mycobacterial sample, the method comprising the steps of:(a) treating the cells to release pre-rRNA, wherein said treatment comprises the steps of:(i) pretreating the cells by enzymatic degradation using both lysozyme and protease until their cell walls are rendered porous to expose their cell membranes making the cells susceptible to lysis by steps (ii) and (iii);(ii) contacting the pretreated cells with a combination of a magnesium chelator, a nonionic detergent and an anionic detergent; and(iii) heating the cells to between about between 75.degree.-99.degree. C. until the mycobacterial cells are lysed, and(b) detecting the pre-rRNA using at least one oligonucleotide probe which is capable of selectively hybridizing, under hybridizing conditions, to a region of the pre-rRNA that is not present in a mature mycobacterial rRNA.