Molecular Methods for Microbial Identification and Typing by K. J. Towner, A. Cockayne (auth.)

By K. J. Towner, A. Cockayne (auth.)

The exact id and typing of microbes is vital for staff energetic in all fields of microbiology. Many examples of contemporary molecular tools were hid in clinical and clinical literature yet this introductory textual content considers the potential functions of such equipment and compares their merits and disadvantages.

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Extra info for Molecular Methods for Microbial Identification and Typing

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2 PULSED-FIELD GEL ELECTROPHORESIS (PFGE) (a) Basic principles A solution to the problems of analysis caused by complex chromosomal fingerprint patterns is to use a 'rare-cutting' restriction endonuclease to generate only a limited number of large DNA fragments. As stated in the previous section, such large fragments cannot be separated readily by conventional agarose gel electrophoresis. The only technique available currently that is capable of separating DNA molecules in the 50 Analysis of chromosomal DNA 45 kb-12 Mb range physically is pulsed-field gel electrophoresis (PFGE), first described by Schwartz and Cantor (1984).

345-76. B. I. (1989) Serotyping Bordetella pertussis strains. Vaccine, 7, 491-4. , Polveroni, G. and Abadie, G. (1988) Serotyping of Pasteurella haemolytica - comparison and adjustment of antigenic extracts and techniques. Revue de Medecine Veterinaire, 139, 719-22. , Nicklon, S. R. (1977) DNA sequencing with chainterminating inhibitors. Proceedings of the National Academy of Sciences of the USA, 74, 5463-7. , Maximescu, P. and Meitert, E. (1979) Corynebacterium diphtheriae. Microbiological methods used in clinical and epidemiological investigations, in Methods in Microbiology, vol.

Such changes may result simply from genetic switching mechanisms rather than real differences between strains. 1), the possibility of additional bands being caused by the presence of extrachromosomal plasmid or bacteriophage DNA should also be considered. Finally, it is important to note that PFGE tends to reflect the gross clonal structure of a collection of strains, rather than the 'fine architecture' which may be of more interest to epidemiologists. This also has the consequence that different 'rare-cutting' enzymes may appear to generate different typing results.

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