METHOD OF DETECTION AND CHARACTERIZATION OF TOXINOGENIC STRAIN CLOSTRIDIUM DIFFICILE Russian patent published in 2017 - IPC C12Q1/68 C12N15/11 C12M1/00 

FIELD: biotechnology.

SUBSTANCE: invention relates to biotechnology. Method of detection and characterization of toxinogenic strain Clostridium difficile is described in sample, in which following steps are made: a. sample is provided; b. in multiplexed PCR-analysis: i) sample is analyzed in relation to presence or absence of tcdB citotoxine gene; ii) sample is analyzed in relation to presence or absence of following deletions in gene tcdC: a) deletions of 18 base pairs in SEQ ID NO: 1 from nucleotide 330 to nucleotide 347; b) deletions of 39 base pairs in SEQ ID NO: 1 as presented in SEQ ID NO: 14; c) deletions of single nucleotide in position 117 SEQ ID NO: 1. Wherein multiplex PCR-amplification is quantitative real time PCR, where sample is additionally analyzed on presence or absence of deletions of 1.8 kbp of tcdA enterotoxin gene, where sample is additionally analyzed with respect to presence or absence of cdtA and/or cdtB binary toxin and wherein a. if sequence of gene tcdB is present, deletion of tcdA is absent, deletion of single nucleotide in position 117 SEQ ID NO: 1, or deletion of 18 BP, or deletion of 39 BP are absent, this test is considered to contain toxinogenic strain Clostridium difficile, b. if sequence of gene tcdB is present, deletion of tcdA is absent, deletion of single nucleotide is present in position 117 SEQ ID NO: 1, deletion of 18 base pairs is present and gene of binary toxin cdtA/B is present, this sample is considered to be of ribotype 027 strain Clostridium difficile, c. if sequence of gene tcdB is present, deletion of tcdA is absent, deletion of single nucleotide in position 117 SEQ ID NO: 1, or deletion of 18 BP, or deletion of 39 base pairs are present in SEQ ID NO: 1 as presented in SEQ ID NO: 14, and there is no gene cdtA/B of binary toxin, this sample is considered to be of ribotype 017 strain Clostridium difficile, and d. if sequence of gene tcdB is present, deletion of tcdA is absent, deletion of 39 base pairs is present in SEQ ID NO: 1 as presented in SEQ ID NO: 14, gene cdtA/B of binary toxin is present, this test is considered to be of ribotype 078 strain Clostridium difficile.

EFFECT: invention extends range of methods of diagnostics.

12 cl, 2 dwg, 1 tbl