FIELD: phthisiobacteriology.
SUBSTANCE: the present innovation deals with laboratory diagnostics of multiple curative tuberculosis. For solving the task it is necessary to detect specific mutations in gene rpoB with polymerase chain reaction with allel-specific primers for codons 516, 526,531. At the absence of mutations in the given codon it is possible to observe a codon-dependent allel-specific PCR-amplification of gene rpoB fragment; availability of mutation in position correspondent to primer's 3'-terminal leads to nonmating of primer and matrix and polymerase's incapacity for the synthesis of complementary fragment. Thus, availability of mutation (RIF-resistant phenotype) leads to the absence of indicator fragment. Simultaneously, it is necessary to conduct three PCR reactions oriented for three codons 516, 526, 531, and PCR result should be analyzed due to electrophoresis in horizontal agarose mini-gel. At the lack of one out of indicator fragments 167, 181, 214 it is possible to conclude upon mutation and mycobacterial resistance to rifampicin.
EFFECT: higher accuracy of quick test detection.
3 dwg, 2 ex
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Authors
Dates
2006-11-20—Published
2003-05-27—Filed