A SEMINESTED POLYMERASE CHAIN REACTION (PCR) METHOD FOR RAPID DETECTION AND SPECIFIC IDENTIFICATION OF MYCOBACTERIUM CHELONAE

摘要

A SEMINESTED POLYMERASE CHAIN REACTION (PCR) METHOD FOR RAPID DETECTION AND SPECIFIC IDENTIFICATION OF MYCOBACTERIUM CHELONAE A seminested Polymerase Chain Reaction (PCR) method of rapid detection and specific identification of mycobacterium chelonae (rapid grower of Mycobacteria) using species-specific primers derived from the internal transcibed spacer (ITS) sequences between 16S and 23 rRNA, of the said mycobacterium, the said primer set comprising the oligonucleotidesd VRF2 5'GACAACACAATGTGTTGCC 3' (complement of 172-190) designed from the available DNA sequence of M.chelonae accession number M15467 of the ITS region for detection of M.chelonae the said primers have two definite sizes of 132 bp (base pair) at the end of the I round of amplification and 94 bp (base pair) at the end of the II round of amplification for easy discrimination thereof by gel electrophoresis.