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Clinical Microbiology Reviews, April 2000, p. 167-195, Vol. 13, No. 2
0893-8512/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Applications of Flow Cytometry to Clinical Microbiologydagger

Alberto Álvarez-Barrientos,1,2 Javier Arroyo,1,3 Rafael Cantón,1,4 César Nombela,1 and Miguel Sánchez-Pérez1,2,*

Departamento de Microbiología II, Facultad de Farmacia,1 Centro de Citometría de Flujo y Microscopía Confocal,2 and Centro de Secuenciación Automatizada de DNA,3 Universidad Complutense de Madrid, and Servicio de Microbiología del Hospital Ramón y Cajal, Carretera Colmenar,4 Madrid, Spain

Classical microbiology techniques are relatively slow in comparison to other analytical techniques, in many cases due to the need to culture the microorganisms. Furthermore, classical approaches are difficult with unculturable microorganisms. More recently, the emergence of molecular biology techniques, particularly those on antibodies and nucleic acid probes combined with amplification techniques, has provided speediness and specificity to microbiological diagnosis. Flow cytometry (FCM) allows single- or multiple-microbe detection in clinical samples in an easy, reliable, and fast way. Microbes can be identified on the basis of their peculiar cytometric parameters or by means of certain fluorochromes that can be used either independently or bound to specific antibodies or oligonucleotides. FCM has permitted the development of quantitative procedures to assess antimicrobial susceptibility and drug cytotoxicity in a rapid, accurate, and highly reproducible way. Furthermore, this technique allows the monitoring of in vitro antimicrobial activity and of antimicrobial treatments ex vivo. The most outstanding contribution of FCM is the possibility of detecting the presence of heterogeneous populations with different responses to antimicrobial treatments. Despite these advantages, the application of FCM in clinical microbiology is not yet widespread, probably due to the lack of access to flow cytometers or the lack of knowledge about the potential of this technique. One of the goals of this review is to attempt to mitigate this latter circumstance. We are convinced that in the near future, the availability of commercial kits should increase the use of this technique in the clinical microbiology laboratory.

* Corresponding author. Present address: Catedratico de Microbiología, Dto. di Microbiología y Genetica, Edificio Departamental, Campus Miguel de Unamuno, Universidad de Salamanca, 37007 Salamanca, Spain. Phone: 34-923 294400. Fax: 34-923 224876. E-mail: misanper{at}gugu.usal.es.

dagger We dedicate this review to Luis Carrasco.

Clinical Microbiology Reviews, April 2000, p. 167-195, Vol. 13, No. 2
0893-8512/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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