Rapid identification of mycobacteria to the species level by polymerase chain reaction and restriction enzyme analysis

TLDR

The authors used PCR of the 65‑kDa protein gene followed by restriction enzyme analysis with BstEII and HaeIII to differentiate 330 mycobacterial isolates to species or subspecies level. The method requires no hybridization or radioactivity and can be completed within one working day.

Abstract

A method for the rapid identification of mycobacteria to the species level was developed on the basis of evaluation by the polymerase chain reaction (PCR) of the gene encoding for the 65-kDa protein. The method involves restriction enzyme analysis of PCR products obtained with primers common to all mycobacteria. Using two restriction enzymes, BstEII and HaeIII, medically relevant and other frequent laboratory isolates were differentiated to the species or subspecies level by PCR-restriction enzyme pattern analysis. PCR-restriction enzyme pattern analysis was performed on isolates (n = 330) from solid and fluid culture media, including BACTEC, or from frozen and lyophilized stocks. The procedure does not involve hybridization steps or the use of radioactivity and can be completed within 1 working day.