Abstract
Consumption, white plague, tuberculosis… Only relatively recently, this disease has ceased to be an absolutely death sentence for infected people, but problems of the spread and diagnosis of the disease are still relevant. This paper presents results of the development of a new loop isothermal amplification (LAMP) assay, named TB-ISATEST, which targeting the species-specific gene rv2341 for the differentiation of Mycobacterium tuberculosis from non-tuberculosis mycobacteria. The assay is applicable for quantitative analysis of genomic DNA and allows detecting tenfold difference in concentration. The results of amplification optimization using a unique two-stage protocol based on the method of orthogonal Taguchi matrices are presented for the first time. A theoretical interpretation of the high amplification efficiency values observed in the LAMP reaction is proposed. Limit of detection of the developed assay is 40 copies of genomic DNA per reaction and amplification requires 15 min. In terms of the combination of characteristics, the TB-ISATEST assay surpasses all the known ways for identifying M. tuberculosis by the LAMP method.
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