SUMMARY 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.

Clinical Microbiology

Microbiological Techniques

10.1128/cmr.13.2.167

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Citations (251)

Methodologies for in vitro and in vivo evaluation of efficacy of antifungal and antibiofilm agents and surface coatings against fungal biofilms

Microbial Cell (2018)

Patrick Van Dijck Jelmer Sjollema Bruno P.A. Cammue Katrien Lagrou Judith Berman

Unlike superficial fungal infections of the skin and nails, which are the most common fungal diseases in humans, invasive fungal infections carry high morbidity and mortality, particularly those associated with biofilm formation on indwelling medical devices. Therapeutic management of these complex diseases is often complicated by the rise in resistance to the commonly used antifungal agents. Therefore, the availability of accurate susceptibility testing methods for determining antifungal resistance, as well as discovery of novel antifungal and antibiofilm agents, are key priorities in medical mycology research. To direct advancements in this field, here we present an overview of the methods currently available for determining (i) the susceptibility or resistance of fungal isolates or biofilms to antifungal or antibiofilm compounds and compound combinations; (ii) the in vivo efficacy of antifungal and antibiofilm compounds and compound combinations; and (iii) the in vitro and in vivo performance of anti-infective coatings and materials to prevent fungal biofilm-based infections.

Antifungal drugs

Echinocandins

10.15698/mic2018.07.638

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Citations (105)

Bacterial infections in cirrhosis: A position statement based on the EASL Special Conference 2013

Journal of Hepatology (2014)

Rajiv Jalan Javier Fernández Reiner Wiest Bernd Schnabl Richard Moreau

Bacterial infections are very common and represent one of the most important reasons of progression of liver failure, development of liver-related complications, and mortality in patients with cirrhosis. In fact, bacterial infections may be a triggering factor for the occurrence of gastrointestinal bleeding, hypervolemic hyponatremia, hepatic encephalopathy, kidney failure, and development of acute-on-chronic liver failure. Moreover, infections are a very common cause of repeated hospitalizations, impaired health-related quality of life, and increased healthcare costs in cirrhosis. Bacterial infections develop as a consequence of immune dysfunction that occurs progressively during the course of cirrhosis. In a significant proportion of patients, infections are caused by gram-negative bacteria from intestinal origin, yet gram-positive bacteria are a frequent cause of infection, particularly in hospitalized patients. In recent years, infections caused by multidrug-resistant bacteria are becoming an important clinical problem in many countries.The reduction of the negative clinical impact of infections in patients with cirrhosis may be achieved by a combination of prophylactic measures, such as administration of antibiotics, to reduce the occurrence of infections in high-risk groups together with early identification and management of infection once it has developed. Investigation on the mechanisms of altered gut microflora, translocation of bacteria, and immune dysfunction may help develop more effective and safe methods of prevention compared to those that are currently available. Moreover, research on biomarkers of early infection may be useful in early diagnosis and treatment of infections.The current manuscript reports an in-depth review and a position statement on bacterial infections in cirrhosis.

Spontaneous bacterial peritonitis

Hepatic Encephalopathy

10.1016/j.jhep.2014.01.024

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Citations (813)

Activity of the new cephalosporin CXA-101 (FR264205) against Pseudomonas aeruginosa isolates from chronically-infected cystic fibrosis patients

Clinical Microbiology and Infection (2010)

Laura Zamorano Carlos Juan A. Fernández-Olmos Y L Ge Rafael Cantón

Cefepime

Tazobactam

Piperacillin/tazobactam

10.1111/j.1469-0691.2010.03130.x

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Citations (38)

Method-Dependent Epidemiological Cutoff Values for Detection of Triazole Resistance in Candida and Aspergillus Species for the Sensititre YeastOne Colorimetric Broth and Etest Agar Diffusion Methods

Antimicrobial Agents and Chemotherapy (2018)

Ana Espinel‐Ingroff John Turnidge Ana Alastruey‐Izquierdo Françoise Botterel Rafael Cantón

Although the Sensititre Yeast-One (SYO) and Etest methods are widely utilized, interpretive criteria are not available for triazole susceptibility testing of Candida or Aspergillus species. We collected fluconazole, itraconazole, posaconazole, and voriconazole SYO and Etest MICs from 39 laboratories representing all continents for (method/agent-dependent) 11,171 Candida albicans , 215 C. dubliniensis , 4,418 C. glabrata species complex, 157 C. guilliermondii ( Meyerozyma guilliermondii ), 676 C. krusei ( Pichia kudriavzevii ), 298 C. lusitaniae ( Clavispora lusitaniae ), 911 C. parapsilosis sensu stricto , 3,691 C. parapsilosis species complex, 36 C. metapsilosis , 110 C. orthopsilosis , 1,854 C. tropicalis , 244 Saccharomyces cerevisiae , 1,409 Aspergillus fumigatus , 389 A. flavus , 130 A. nidulans , 233 A. niger , and 302 A. terreus complex isolates.

Etest

Candida parapsilosis

Posaconazole

Candida glabrata

Candida krusei

10.1128/aac.01651-18

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Citations (64)

Holistic medicine: reactivation of an old aspiration.

PubMed (1980)

Rafael Cantón

Source

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Citations (3)

Improvement of intestinal comfort in cystic fibrosis patients after probiotics consumption

Journal of Cystic Fibrosis (2009)

Rosa del Campo María Garriga Jorge Agrimbau A. Lamas L. Máiz

10.1016/s1569-1993(09)60346-5

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Citations (3)

The E-Test as an epidemiologic tool

Diagnostic Microbiology and Infectious Disease (1992)

Fernando Baquero Rafael Cantón J Martínez-Beltrán Anne Bolmström

10.1016/0732-8893(92)90095-b

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Citations (18)

Wild-Type MIC Distributions and Epidemiological Cutoff Values for Amphotericin B, Flucytosine, and Itraconazole and Candida spp. as Determined by CLSI Broth Microdilution

Journal of Clinical Microbiology (2012)

Michael A. Pfaller Ana Espinel-Ingroff Rafael Cantón Mariana Castanheira M. Cuenca‐Estrella

Clinical breakpoints (CBPs) and epidemiological cutoff values (ECVs) have been established for several Candida spp. and the newer triazoles and echinocandins but are not yet available for older antifungal agents, such as amphotericin B, flucytosine, or itraconazole. We determined species-specific ECVs for amphotericin B (AMB), flucytosine (FC) and itraconazole (ITR) for eight Candida spp. (30,221 strains) using isolates from 16 different laboratories in Brazil, Canada, Europe, and the United States, all tested by the CLSI reference microdilution method. The calculated 24- and 48-h ECVs expressed in μg/ml (and the percentages of isolates that had MICs less than or equal to the ECV) for AMB, FC, and ITR, respectively, were 2 (99.8)/2 (99.2), 0.5 (94.2)/1 (91.4), and 0.12 (95.0)/0.12 (92.9) for C. albicans; 2 (99.6)/2 (98.7), 0.5 (98.0)/0.5 (97.5), and 2 (95.2)/4 (93.5) for C. glabrata; 2 (99.7)/2 (97.3), 0.5 (98.7)/0.5 (97.8), and 05. (99.7)/0.5 (98.5) for C. parapsilosis; 2 (99.8)/2 (99.2), 0.5 (93.0)/1 (90.5), and 0.5 (97.8)/0.5 (93.9) for C. tropicalis; 2 (99.3)/4 (100.0), 32 (99.4)/32 (99.3), and 1 (99.0)/2 (100.0) for C. krusei; 2 (100.0)/4 (100.0), 0.5 (95.3)/1 (92.9), and 0.5 (95.8)/0.5 (98.1) for C. lusitaniae; -/2 (100.0), 0.5 (98.8)/0.5 (97.7), and 0.25 (97.6)/0.25 (96.9) for C. dubliniensis; and 2 (100.0)/2 (100.0), 1 (92.7)/-, and 1 (100.0)/2 (100.0) for C. guilliermondii. In the absence of species-specific CBP values, these wild-type (WT) MIC distributions and ECVs will be useful for monitoring the emergence of reduced susceptibility to these well-established antifungal agents.

Flucytosine

Broth microdilution

Candida parapsilosis

Candida glabrata

Candida krusei

10.1128/jcm.00248-12

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Citations (157)

[In vitro activity of posaconazole against yeasts isolated in blood cultures].

PubMed (2002)

Rafael Cantón Javier Pemán A Orero A Viudes Gil J

The in vitro activity of posaconazole against Candida species isolated from blood cultures and the influence of incubation time was studied and compared with that of fluconazole. A total of 112 isolates were studied: 32 Candida albicans, 33 C. parapsilosis, 17 C. tropicalis, 8 C. glabrata, 8 C. guilliermondii, 3 C. famata, 2 C. lusitaniae, 1 C. lipolytica, 1 C. inconspicua, 1 C. lambica, 3 Saccharomyces cerevisiae, 1 Blastoschizomyces capitatus, 1 Geotricum spp. and 1 Pichia omheri. The MIC was obtained using the M27-A microdilution method described by the NCCLS for Candida spp. and Cryptococcus neoformans. The species most susceptible to posaconazole were C. parapsilosis (MCI90 0.016 mg/l), C. guilliermondii (MIC90 0.12 mg/l), C. glabrata (MCI90 0.5 mg/l) and Candida spp. (MCI90 0.25 mg/l). However, this azole did not improve the activity of fluconazole against C. tropicalis (MIC90 8 mg/l) and C. albicans (MCI90 mg/l). The time of reading was important in the detection of resistance, as the number of strains resistant to fluconazole or posaconazole was higher at 48 hours than at 24 hours for C. albicans and C. tropicalis. All the other species of Candida were susceptible at both reading times.

Posaconazole

Candida parapsilosis

Candida glabrata

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Citations (5)