PMID- 7751374
OWN - NLM
STAT- MEDLINE
DCOM- 19950622
LR  - 20210526
IS  - 0095-1137 (Print)
IS  - 1098-660X (Electronic)
IS  - 0095-1137 (Linking)
VI  - 33
IP  - 3
DP  - 1995 Mar
TI  - Susceptibility testing of Candida albicans and Aspergillus species by a simple 
      microtiter menadione-augmented 
      3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide assay.
PG  - 661-7
AB  - We describe a simple microtiter method for determining the susceptibility of 
      Candida albicans and hyphal forms of Aspergillus fumigatus against antifungal 
      agents. The assay measures mitochondrial respiration by determining reduction of 
      3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) to 
      formazan, a process that is enhanced in the presence of menadione. C. albicans or 
      conidial suspensions of A. fumigatus are seeded into microtiter plates. Hyphal 
      outgrowth of Aspergillus spp. was achieved by a 12 to 14-h culture at 30 degrees 
      C. Antifungal agents (amphotericin B, fluconazole, itraconazole) were added to 
      the cultures for 24 h. Thereafter, incubations were continued for 3 h in the 
      presence of MTT plus 0.1 mM menadione. Formazan formation was quantified 
      photometrically after extraction of the formazan with acid isopropanol. 
      Well-defined dose-response curves reflecting impairment of mitochondrial function 
      by the antifungal agents were obtained. With C. albicans, the results correlated 
      excellently with the MIC determinations performed according to the standard 
      macrodilution procedure. In confirmation of a recent report, it was found that 
      fluconazole was unable to exert its fungistatic action on a sensitive C. albicans 
      strain in the presence of serum. The presented method can easily be integrated in 
      the standard repertoire of a diagnostic microbiology laboratory and should prove 
      useful as a means to assess the antifungal action of various agents on yeasts and 
      filamentous fungi in the presence and absence of serum proteins or body fluids.
FAU - Jahn, B
AU  - Jahn B
AD  - Institute of Medical Microbiology and Hygiene, Hochhaus am Augustusplatz, Mainz, 
      Germany.
FAU - Martin, E
AU  - Martin E
FAU - Stueben, A
AU  - Stueben A
FAU - Bhakdi, S
AU  - Bhakdi S
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - J Clin Microbiol
JT  - Journal of clinical microbiology
JID - 7505564
RN  - 0 (Formazans)
RN  - 0 (Tetrazolium Salts)
RN  - 0 (Thiazoles)
RN  - 117038-70-7 
      (2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-5-((phenylamino)carbonyl)-2H-tetrazolium 
      hydroxide)
RN  - 12001-79-5 (Vitamin K)
RN  - 304NUG5GF4 (Itraconazole)
RN  - 7XU7A7DROE (Amphotericin B)
RN  - 8VZV102JFY (Fluconazole)
RN  - EUY85H477I (thiazolyl blue)
SB  - IM
MH  - Amphotericin B/pharmacology
MH  - Aspergillus fumigatus/*drug effects/metabolism
MH  - Candida albicans/*drug effects/metabolism
MH  - Dose-Response Relationship, Drug
MH  - Fluconazole/pharmacology
MH  - Formazans/analysis
MH  - Itraconazole/pharmacology
MH  - Microbial Sensitivity Tests/*methods
MH  - Mitochondria/metabolism
MH  - Tetrazolium Salts/*metabolism
MH  - Thiazoles/*metabolism
MH  - Vitamin K/*pharmacology
PMC - PMC228010
EDAT- 1995/03/01 00:00
MHDA- 1995/03/01 00:01
PMCR- 1995/03/01
CRDT- 1995/03/01 00:00
PHST- 1995/03/01 00:00 [pubmed]
PHST- 1995/03/01 00:01 [medline]
PHST- 1995/03/01 00:00 [entrez]
PHST- 1995/03/01 00:00 [pmc-release]
AID - 10.1128/jcm.33.3.661-667.1995 [doi]
PST - ppublish
SO  - J Clin Microbiol. 1995 Mar;33(3):661-7. doi: 10.1128/jcm.33.3.661-667.1995.