The MTB/MDR Test Kit(Sanity 2.0) (Abbreviation: Sanity-2 MTB/MDR) is a cartridge-based test kit for qualitative detection of Mycobacterium tuberculosis complex (MTBC) and multidrug-resistance mutations (including rifampin-resistant mutations and isoniazid-resistant mutations) in Mycobacterium tuberculosis (MTB) from sputum samples. This kit is designed to run on Sanity 2.0, which integrates nucleic acid extraction, real-time polymerase chain reaction (rt-PCR), analysis of melting curve and the result interpretation into a fully automatic instrument (all-in-one machine).
The Sanity-2 MTB/MDR uses Taqman probe-based real-time polymerase chain reaction (rt-PCR) to qualitatively detect IS6110 gene that is specific for MTBC as well as a non-human internal control (Arabidopsis thaliana SUC2 gene).
At the same time, the Sanity-2 MTB/MDR employs dually labeled probes to target the excess single-stranded amplicons generated by asymmetric PCR; then uses melting curve analysis to generate the minus derivative of the fluorescence intensity versus temperature, and compares the melting temperature (Tm) to gain mutation information of the sequences. Where, perfect match of target sequence and probe gives the highest Tm; partially mismatch, e.g., point mutation, insertion or deletion, would cause decreased Tm than that of perfect match. The decreased degree is related to the type of point mutation, base number of insertion or deletion as well as the mutation location. The Sanity-2 MTB/MDR covers the mutations of rpoB (codons 507~533) to determine the resistance of rifampin; the mutations of ahpC promoter region (positions -44~-30 and -15~4), inhA promoter region (positions -17~-8), and katG codon 315, to determine the resistance of isoniazid.
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