Antifungal susceptibility testing

Rex, John H.; Pfaller, Michael A.; Rinaldi, Michael G.; Polak, Annemarie; Galgiani, John N.

doi:10.1128/cmr.6.4.367

Cited by 278 publications

(214 citation statements)

References 128 publications

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“…The lid was left ajar for 40 min to allow the acetone to evaporate. Preliminary tests showed that a 40 min drying time was optimal for minimizing variation ( Variation in fungal growth rates among assays was minimized by using a stock spore suspension (Rex et al 1993). The stock suspension was made with spores of Aspergillus sydowii isolated from San Salvador, Bahamas (Smith et al 1996) and filtered through gauze to remove hyphal mats and then suspended in de-ionized water at a concentration of 2 150 000 spores ml -1 .…”

Section: Methodsmentioning

confidence: 99%

Size structure and geographic variation in chemical resistance of sea fan corals Gorgonia ventalina to a fungal pathogen

Dube¹,

Kim²,

Alker³

et al. 2002

Mar. Ecol. Prog. Ser.

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Section: Methodsmentioning

confidence: 99%

Size structure and geographic variation in chemical resistance of sea fan corals Gorgonia ventalina to a fungal pathogen

Dube¹,

Kim²,

Alker³

et al. 2002

Mar. Ecol. Prog. Ser.

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“…The test method can affect the level of antifungal activity determined (Rex et al, 1993(Rex et al, , 1995. The NCCLS recommend the use of RPMI 1640 buffered to pH 7 .…”

Section: Effect Of Test Methods On In Vitro Activity Of Dihydropyrrolementioning

confidence: 99%

In vitro antifungal activity of 2-(3,4-dimethyl-2,5-dihydro-1H-pyrrol-2-yl)-1-methylethyl pentanoate, a dihydropyrrole derivative

Dabur

Chhillar

Yadav

et al. 2005

Journal of Medical Microbiology

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A novel compound 2-(3,4-dimethyl-2,5-dihydro-1H-pyrrol-2-yl)-1-methylethyl pentanoate was isolated from the plant Datura metel L. The in vitro activity of this dihydropyrrole derivative against Aspergillus and Candida species was evaluated by using standard methods approved by the National Committee for Clinical Laboratory Standards. The compound was found to be active against all the species tested, namely Candida albicans, Candida tropicalis, Aspergillus fumigatus, Aspergillus flavus and Aspergillus niger. The MIC at which more than 90 % of growth was inhibited (MIC 90 ) by the compound ranged from 21 . 87 to 43 . 75 ìg ml À1 against various fungal species by microbroth dilution assay. Since the compound 2-(3,4-dimethyl-2,5-dihydro-1H-pyrrol-2-yl)-1-methylethyl pentanoate has antifungal activity it can be explored further to develop new antimycotic drugs. INTRODUCTIONPathogens belonging to fungal genera Aspergillus and Candida are ubiquitous and global in distribution (Randhawa & Khan, 1987). These infections are increasingly recognized as an emerging threat to public health. In immunocompromised hosts, infections by Aspergillus, Candida, Histoplasma, etc., become invasive and disseminate from the primary site of infection to other parts of the body including the gut, kidney, spleen and brain.Invasive aspergillosis is reported to be associated with a mortality rate of 55 % (Denning & Stevens, 1990). Mortality due to Aspergillus infection in bone marrow transplant recipients was observed to be as high as 80 % despite appropriate chemotherapy (Meyer, 1990). Cerebral aspergillosis presents the symptoms of acute meningitis and is always fatal.Candida species have been found to be the fourth most prevalent group of pathogens and have been isolated from 8 % of patients with nosocomial bloodstream infections (Pfaller, 1994). They are also identified as critical pathogens in infections of wounds and other body fluids (Powderly et al., 1988). The change in mortality rate associated with disseminated candidiasis has been insignificant even after treatment with effective antifungal drugs (Pfaller et al., 1999).The drugs currently available for treatment of various fungal infections are primarily polyenes and the azole class of compounds. Amphotericin B, which is considered to be the drug of choice, has been found to be highly nephrotoxic and less effective in invasive aspergillosis (Powderly et al., 1988;Rex et al., 1995). Further, the development of fungal resistance against most of the available drugs has been observed (Pfaller et al., 1998a, b;Powderly, 1994). The increased occurrence of mycoses in immunocompromised patients and the development of resistance in fungi to current drugs has emphasized the need for developing new antifungal compounds with minimal adverse effects in humans.Several bioactive molecules, including antifungals, from plants have been reported. 2-(3,4-dimethyl-2,5-dihydro-1H-pyrrol-2-yl)-1-methylethyl pentanoate, a novel antifungal molecule, was isolated from the plant Datura metel L (Rajesh et...

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“…Despite great advances in the standardization of antifungal susceptibility testing, azole endpoint determination continues to be problematic and subjective and a major source of interlaboratory variability (6,18,23). The trailing-growth phenomenon is often responsible for these difficulties, whereby partial inhibition of fungal growth occurs over the range of azole concentrations (6,13,14,23).…”

mentioning

confidence: 99%

“…The trailing-growth phenomenon is often responsible for these difficulties, whereby partial inhibition of fungal growth occurs over the range of azole concentrations (6,13,14,23).…”

mentioning

confidence: 99%

Novel Fluorescent Broth Microdilution Method for Fluconazole Susceptibility Testing of Candida albicans

2001

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A comparative evaluation of the reference National Committee for Clinical Laboratory Standards (NCCLS) broth microdilution method with a novel fluorescent carboxyfluorescein diacetate (CFDA)-modified microdilution method for the susceptibility testing of fluconazole was conducted with 68 Candida strains, including 53 Candida albicans, 5 Candida tropicalis, 5 Candida glabrata, and 5 Candida parapsilosis strains. We found trailing endpoints and discordant fluconazole MICs of <8 g/ml at 24 h and of >64 g/ml at 48 h for 12 of the C. albicans strains. These strains satisfy the definition of the low-high MIC phenotype. All 12 low-high phenotype strains were correctly shown to be susceptible at 48 h with the CFDA-modified microdilution method. For the 41 non-low-high phenotype C. albicans strains, the CFDA-modified microdilution method yielded 97.6% (40 of 41 strains) agreement within ؎1 dilution at 24 h compared with the reference method and 92.7% (38 of 41 strains) agreement within ؎1 dilution at 48 h compared with the reference method. The five strains each from C. tropicalis, C. glabrata, and C. parapsilosis that were tested showed 100% agreement within ؎2 dilutions for the two methods being evaluated.Candida is the fourth most common cause of nosocomial bloodstream infection in the United States (2, 4), and the rate of primary bloodstream infections continue to increase (3, 9). In the Americas, Candida albicans is the species most frequently isolated from the bloodstream (16). Candidemia is often difficult to diagnose and refractory to therapy. The attributable mortality rate is 38% in the United States (28) and between 19 and 23% in Canada (9,25,29).Use of the broad-spectrum antifungal fluconazole for the treatment of serious systemic Candida infections has increased. Fluconazole is a less toxic alternative to amphotericin B and has been recommended as primary therapy for candidemia in nonneutropenic and stable neutropenic patients who have not received prior fluconazole treatment and in whom Candida krusei is unlikely (5, 24). In vitro susceptibility testing for fluconazole is of clinical importance, as therapeutic success depends substantially on achieving plasma fluconazole levels that are sufficiently higher than MICs (22).Despite great advances in the standardization of antifungal susceptibility testing, azole endpoint determination continues to be problematic and subjective and a major source of interlaboratory variability (6, 18, 23). The trailing-growth phenomenon is often responsible for these difficulties, whereby partial inhibition of fungal growth occurs over the range of azole concentrations (6,13,14,23).Previous work has demonstrated the utility of using fluorescent dyes to assess the viability of C. albicans exposed to amphotericin B (10). We have investigated the use of the vitalityspecific dye carboxyfluorescein diacetate (CFDA) in the standardized NCCLS M27-A broth microdilution method (12) to assess the susceptibility of Candida spp. to fluconazole. In this study we compared the NCCLS microdilu...

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Antifungal susceptibility testing

Cited by 278 publications

References 128 publications

Size structure and geographic variation in chemical resistance of sea fan corals Gorgonia ventalina to a fungal pathogen

Size structure and geographic variation in chemical resistance of sea fan corals Gorgonia ventalina to a fungal pathogen

In vitro antifungal activity of 2-(3,4-dimethyl-2,5-dihydro-1H-pyrrol-2-yl)-1-methylethyl pentanoate, a dihydropyrrole derivative

Novel Fluorescent Broth Microdilution Method for Fluconazole Susceptibility Testing of Candida albicans

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