SIMULTANEOUS DETERMINATION, IDENTIFICATION AND DIFFERENTIATION OF EUBACTERIAL TAXONS USING HYBRIDIZATION ANALYSIS Russian patent published in 2000 - IPC

FIELD: medicine, microbiology, molecular biology. SUBSTANCE: invention relates to determination and identification of at least a single microorganism or simultaneous determination of different microorganisms in a sample. Method involves: (i) release, isolation and/or concentration of polynucleic acids in the sample; (ii) amplification of 16S-23S tRNA spacer region or its part using at least one suitable pair of primers if necessary; (iii) hybridization of polynucleic acids obtained in stages (i) or (ii) by at least with a single and, preferably, more spacer-probe given in Table 1a or its equivalent under the similar conditions of hybridization and washing out, and/or with taxon-specific probe of any spacer sequences given in Fig. 1-103 under the same conditions of hybridization and washing out; (iv) detection of hybrids formed in stage (iii) with each probe used under corresponding conditions of hybridization and washing out; (v) identification of microorganism(s) in the sample based on difference hybridization signals obtained in stage (iv). Invention involves also probe, composition, primer and set for determination and identification of microorganisms. EFFECT: enhanced specificity and sensitivity of assay and identification of microorganisms. 54 cl, 11 tbl, 103 dwg, 9 ex