The aim of this study, was to design and validate 16S rRNA targeted oligonucleotide genus specific primers for amplifying the predominant members of gut flora using polymerase chain reaction. Primers were validated against human faecal samples. Gut flora of a normal individual was compared with that of two diseased individuals. Our observations showed that the genera Lactobacillus, Bacteroides, Peptococcus, Bifidobacterium, and E. coli were invariably present in all studied subjects however, the absence of butyrate producing bacteria Ruminococcus and Peptostreptococcus were significant. Presence of the members of the genus, Campylobacter in both the diseased samples were also unusual.