Ribotyping is a molecular technique for bacterial identification that uses information from rRNA-based phylogenetic analyses. It is rapid and specific method widely used in clinical diagnostics and analysis of microbial communities in food, water, and beverages.
11. Selection of restriction endonuclease by sequence analysis(in silico)
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Ribotyping prototype : Haemophilus influenzae strain Rd
In silico analysis
The presence of 6 ribosomal operon
12. Ribotyping protocol for Haemophilus
influenzae strain Rd
Stage 1-search the conserved restriction endonuclease site within the six
ribosomal operon.
Stage 2-choose the ideal restriction enzyme
Stage3-confirm the conservation of restriction site by the all publicity
available for 16-23s-rRNA sequencing
Stage4-cleavage electrophoresis southern blot hybridization with
a labelled probe containing conserved ribosomal sequences.
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13. Cont.
0 The importance of this method :strain differentiation within the species
0 It implemented for several other bacterial species,eg : S.aureus
E.coli
p.aeroginosa
B.cepacia
N.meningitidis
B.pertussis
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14. Automated ribotyping
0 Was first introduced by dupont qualicon-1995….. Riboprint pattern.
0 This system is reproducible ,convenient and fast
0 A single colony of bacteria is picked up and suspended in lysing buffer
and transferred to the riboprinter in the riboprinter like
conventional ribotyping….
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16. Advantages and disadvantages
Advantages of ribotyping as an identification method:
0 Ribotyping is a fully automated procedure
0 Procedure involves less labor and is standardized
Disadvantages of ribotyping:
0 Expensive because of the equipment used, therefore usually
only performed in reference laboratories
22. references
• Grimont, F., and P. A. D. Grimont. 1986. Ribosomal ribonucleic acid
gene restriction patterns aspotential taxonomic
tools. Ann. Inst. Pasteur Micro- biol.
• Altwegg, M., F. W. Hickman-Brenner, and J. J. Farmer III. 1989.
Ribosomal RNA gene restriction patterns provide increased sensitivity
for typing Salmonella typhi strains. J. Infect. Dis. 160:145–149.
• Bingen, E. H., P. Mariani-Kurkdjian, N. Y. Lambert-Zechovsky, P.
Dejardins, E. Denamur, Y. Aujard, E. Vilmer, and J. Elion. 1992.
Ribotyping provides efficient differentiation of nosocomial Serratia
marcescens isolates in a pediatric hospital. J. Clin. Microbiol.
30:2088–2091.
23. Cont.
Farmer, J. J., III. 1972. Epidemiological differentiation of Serratia marcescens:
typing by bacteriocin sensitivity. Appl. Microbiol. 23:226–231.
• Animal Biotechnology: Models in Discovery and Translationedited by
Ashish Verma, Anchal Singh
