Molecular Susceptibility Testing for <i>Mycobacterium tuberculosis</i>

Abstract Pyrosequencing (PSQ) technology is a rapid real‐time method for sequencing small segments of genomic deoxyribonucleic acid (DNA), often referred to as nucleic acid sequencing‐by‐synthesis. PSQ can be utilized for the molecular detection of mutations associated with drug resistance for Mycobacterium tuberculosis complex (MTBC). PSQ, when utilized directly on clinical specimens, has been shown to shorten the time to treatment of multidrug‐resistant tuberculosis (MDR‐TB) to as much as 5 weeks earlier and has influenced clinicians' decisions to devise effective TB drug regimens. The technology utilizes polymerase chain reaction products amplified from primary clinical samples or cultures to yield single‐stranded DNA to which a sequencing primer can be annealed. Sequences derived from pyrograms are compared to a library of known sequences to determine if the sample has mutations and resistance is predicted or if it has no mutations and susceptibility is suggested.