Advanced Techniques in Diagnostic Microbiology

154 F. Wu and P. Della-Latta
Summary
PFGE is widely used for genotypic characterization of microorganisms. It has
been considered the method of choice for the analysis of most bacterial pathogens
because of its high discriminatory power and reproducibility. Although PFGE is
an important tool for outbreak investigations, it is not indicated for population
analysis of microorganisms. In addition, PFGE is marginally valuable in epidemiological
settings of long-term duration encompassing years or decades or at a
country to continent level (Blanc et al., 2002). Its limitations also include substantial
measurements of time and resources, and the non-typeability of some strains
in some situations. Therefore, a combination of PFGE and PCR-based techniques
may be necessary for genotyping a wide range of microorganisms under these
circumstances.
References
Aires de Sousa, M., & de Lencastre, H. (2004). Bridges from hospitals to the laboratory:
genetic portraits of methicillin-resistant Staphylococcus aureus clones. FEMS Immunol
Med Microbiol, 40, 101–111.
Arbeit, R.D., Arthur, M., Dunn, R., Kim, C., Selander, R.K., & Goldstein, R. (1990). Resolution
of recent evolutionary divergence among Escherichia coli from related lineages:
the application of pulsed field electrophoresis to molecular epidemiology. J Infect Dis,
161, 230–235.
Baba, T., Takeuchi, F., Kuroda, M., Yuzawa, H., Aoki, K., Oguchi, A., Nagai, Y., Iwama,
N., Asano, K., Naimi, T., Kuroda, H., Cui, L., Yamamoto, K., & Hiramatsu, K. (2002).
Genome and virulence determinants of high virulence community-acquired MRSA.
Lancet, 25, 1819–1827.
Bennett, J.E., Izumikawa, K., & Marr, K.A. (2004). Mechanism of increased fluconazole
resistance in Candida glabrata during prophylaxis. Antimicrob Agents Chemother, 48,
1773–1777.
Blanc, D.S., Francioli, P., & Hauser, P.M. (2002). Poor value of pulsed-field gel electrophoresis
to investigate long-term scale epidemiology of methicillin-resistant Staphylococcus
aureus. Infect Genet Evolution, 2, 145–148.
Cantor, C.R., Gaal, A., & Smith, C.L. (1988). High-resolution separation and accurate size
determination in pulsed-field gel electrophoresis of DNA. 3. Effect of electrical field
shape. Biochemistry, 27, 9216–9221.
Carle, G.F., Frank, M., & Olson, M.V. (1986). Electrophoretic separations of large DNA
molecules by periodic inversion of the electric field. Science, 232, 65–68.
Carle, G.F., & Olson, M.V. (1984). Separation of chromosomal DNA molecules from
yeast by orthogonal-field-alteration gel electrophoresis. Nucleic Acids Res, 12, 5647–
5664.
Centers for Disease Control and Prevention. (1999). Four pediatric deaths from communityacquired
methicillin-resistant Staphylococcus aureus—Minnesota and North Dakota,
1997–1999. Morbid Mortal Weekly Rep, 48, 707–710.
Centers for Disease Control and Prevention. (2004). Day care-related outbreaks of
rhamnose-negative Shigella sonnei—six states, June 2001-March 2003. Morbid Mortal
Weekly Rep, 53, 60–63.