National Academy of Agricultural Sciences (NAAS)
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PRINT ISSN : 2319-7692 Online ISSN : 2319-7706 Issues : 12 per year Publisher : Excellent Publishers Email : editorijcmas@gmail.com / submit@ijcmas.com Editor-in-chief: Dr.M.Prakash Index Copernicus ICV 2018: 95.39 NAAS RATING 2020: 5.38 |
The 16S rRNA gene is composed of highly conserved, specie-specific sequences between different species of bacteria and the application of restriction endonucleases on the amplified 16S rRNA gene is a novel diagnostic tool in molecular characterization of bacterial isolates. In a study aimed at characterizing bacteria using some selected endonucleases, a total of 31 isolates collected from the stock culture in the Department of Medical Laboratory Science were re-identified using standard microbiological procedures as Pseudomonas aeruginosa, Klebsiella pneumoniae, Escherichia coli and Alcaligenes faecalis. The isolates were subjected to antibiotic susceptibility testing to ascertain their susceptibility pattern using the agar disc diffusion method; followed by DNA extraction, purification, amplification, gel electrophoresis and sequencing. The Sequences were edited using trace edit, Blasted and Restriction site annotation was carried out using Geneious. Antibiotic susceptibility testing results showed that K. pneumoniae is susceptible to Streptomycin, Ciprofloxacin, Gentamicin, Nalidixic Acid, Pefloxacin, Septrin, Ofloxacin and resistant to Augmentin, Ampicillin, Cephalosporin. Restriction site annotation of the sequences revealed that Xma1 cuts Klebsiella pneumoniae and Pseudomonas spp. at position 499 while EcoR1cuts both Klebsiella pneumoniae and Bacillus spp. at position 561; showing that both Xma1 and EcoR1 cannot be used as restriction markers to differentiate between Klebsiella pneumoniae and Pseudomonas spp. and Bacillus spp. respectively.
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