The MDR-TB Test Kit (MMCA) (Abbreviation: MDR-TB Test Kit) is an in vitro diagnostic reagent used for qualitative detection of multidrug-resistance mutations (rifampin-resistant mutations and isoniazid-resistant mutations) in Mycobacterium tuberculosis from cultured Mycobacterium tuberculosis specimens or sputum samples. The kit employs PCR and multicolor melting curve analysis (MMCA) methods, and targets 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.
The MDR-TB Test Kit 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.
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