CHARACTERIZATION AND CLASSIFICATION OF AEROMONAS SPP. BY REP -PCR, BOX – PCR, ERIC – PCR GENOMIC FINGERPRINTING AND MLST ANALYSIS
ABSTRACT
Aeromonas spp. are Gram-negative, non-spore-forming, rod-shaped bacteria. They widely distributed and complex group of ubiquitous bacteria. However historically the Aeromonas genus belongs in the family Vibrionaceae, but later placed in family, the Aeromonadaceae. They are also often isolated from clinical (Krovacek et al. 1994; Carnahan and Altwegg 1996; Kuhn et al. 1997), environmental (Araujo et al. 1990; Burke et al. 1984; Krovacec et al. 1994), and food samples (Buchanan and Palumbo 1985; Abeyta et al. 1986; Abeyta and Wekell 1988). Members of the genus are important pathogens of fish and a variety of animals. In humans they cause opportunistic infections and gastroenteritis, but their role in gastroenteritis is unclear. In spite of a significant number of virulence determinants produced by Aeromonas spp. their association with gastroenteritis is not clearly linked. Different factors have been described including the cytolytic enterotoxin gene AHCYTOEN and aerolysin genes (Kingombe et al. 1999), and haemolysin genes from A. hydrophila, and A. sobria (Hirono et al. 1992), A. salmonicida (Hirono and Aoki 1993) and A. caviae (Wang et al. 1996).
The purpose of this study have been made to compare the presence of different virulence genes and to characterization and classification of microbes by using different molecular techniques, such as polymerase chain reaction (PCR), Randomly amplified polymorphic DNA (RAPD), Restriction fragment length polymorphism (RFLP), Ribotyping, Pulsed Field Gel Electrophoresis (PFGE), Multi Locus Sequence Alignment Analysis (MLST) etc. are now commonly used to identify and characterize this pathogen and We also try to identify the presence of different virulence genes by PCR.. In this study, following molecular techniques are used for bacterial typing. Three categories of conserved repetitive sequences, REP-PCR, BOX-PCR, Enterobacterial Repetitive Intergenic Consensus sequence PCR (ERIC-PCR) and Computer-assisted Pattern Analysis are now being used for characterization and classification of microbes. These studies confirm the presence of different virulent Aeromonas strains by the above techniques. These techniques have wide scope of application in characterization and typing of this pathogen.
Agniswar Sarkar1, 2, Mousumi Saha1 and S. S. Mishra1
1Biotechnology laboratory, Central Inland Fisheries Research Institute (ICAR), Barrackpore, West Bengal ? 700 120, India
2 Dept. of Biotechnology, University of Burdwan, Burdwan, West Bengal-713 104, India
