Detection and identification of genetic diversity among fish pathogens using 16X-rRNA primers
2012
Verma, R.K. | Yadav, P.K. | Gahlawat, S.K.
Many bacterial pathogens can now be detected in samples of various kinds without the need to first culture the organism and phylogenetic relationships by using PCR-based methods. PCR-based genotyping technologies are simple and fast. Under the present investigation, the amplified gel electrophorized product (approx. 500 bp), stained with Ethidium bromide was visualized under U.V. light in gel documentation system. The amplified 16S rDNA was digested with two restriction enzymes SauIIIA and Hinfl at 37°C for 2 h to generate amplified ribosomal DNA restriction analysis (ARDRA) profiles, both the restriction enzymes generated 2-3 fragments in bacterial isolates; these fragments were scored by comparison to a low molecular DNA Ladder (50 bp) and analyzed by 1/0 clustering method of the NT¬SYSpc2.0 programme then a dendrogram displaying hierarchical associations among all isolates was generated. On the basis of similarity coefficient the isolates were divided in five major groups (A, B, C, D and E). Group A and E were found most divergent which have genotypes S. aureus and Streptococcus grp. Ql, respectively. The genotypes KI. oxytoca and A. hydrophila has 83; similarity in group B, whereas P. flourescens and P. aeruginosa have 99; similarity in group D.
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