Patent ID: 7638309
Filing Date: 2009-12-29
Classification: C07H,C12Q

Abstract:
1. A method for detecting pathogenic mycobacteria in a clinical specimen, said method comprising the steps of: (a) clarifying the clinical specimen from contaminant by conventional methods, (b) treating the processed clinical specimen obtained in step (a) with a lysis buffer comprising guanidinium isothiocyanate to inactivate live pathogenic mycobacteria to make the process safe for the user, (c) extracting genomic DNA from the processed clinical specimen obtained from step (b) using a buffer comprising sodium chloride to increase the yield and quality of DNA, (d) designing and synthesizing a set of specific oligonucleotide primers of SEQ ID NO: 5, which is the forward primer and SEQ ID NO: 6, which is the reverse primer, (e) PCR amplifying the genomic DNA from the processed clinical specimen to amplify the DNA stretch having SEQ ID NO: 4 using the specific oligonucleotide primers having SEQ ID NO: 5 and SEQ ID NO: 6, and (f) analyzing the amplified PCR product by restriction fragment length polymorphism (RFLP) analysis for differentiation of the species of the pathogenic mycobacterium for a quick assessment of HIV co-infection.