The Mycobacteria Identification Kit (MMCA) (Abbreviation: Mycobacteria Identification Kit) is an in vitro diagnostic reagent used for mycobacteria identification from cultured mycobacteria specimens or sputum samples. The kit employs PCR and multicolor melting curve analysis (MMCA) methods, and targets the intergenic transcribed spacer (ITS) region between the 16S rRNA and 23S rRNA genes to efficiently detect 51 mycobacteria (Annex table 1 ）, among which, 19 clinically relevant mycobacteria can be identified to the species level, including:
The Mycobacteria Identification 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 according to the specific melting temperature (Tm) to gain the information of the sequence. In the case of the presence of a specific target, a melting peak will arise at a corresponding position, and the Tm value will associate with a specific mycobacteria. Moreover, a fragment of the Arabidopsis thaliana sucrose-proton symporter 2 (SUC2) gene was included as an internal positive control (IPC) for indication of PCR inhibition.
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