The MTB/SL Test Kit (MMCA) (Abbreviation: MTB/SL Test Kit) is an in vitro diagnostic reagent used for qualitative detection of second-line injectable drug-resistance 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 rrs gene (positions 1401, 1402 and 1484) and eis promoter region (positions -37, -14 and -10) to determine the resistance of Second-line injectable drug.
Principle
The MTB/SL 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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