Usefulness of self ligation mediated polymerase chain reaction: a rapid method for fingerprinting in molecular epidemiology of tuberculosis

Abstract

Restriction fragment length polymorphism (RFLP) analysis based on the insertion sequence IS 6110 has been used as one of the powerful tools for epidemiological study of tuberculosis. However this technique requires more than 1 micro-gram of DNA and two days for completion. To overcome these inconvenience, we have modified a PCR-based method, self ligation mediated PCR (SL-PCR) on the molecular epidemiological study. This method uses a pair of primers whose orientations are from inside to outside of IS 6110. The DNA fragments flanking IS 6110 are amplified by the PCR by using the Sau 3A I digested and ligated chromosomal DNA of Mycobacterium tuberculosis strains. By using this method, M. tuberculosis strains can be differentiated within 8 hours.