Journal of Chemotherapy Vol. 17 - n. 4 (361-366) - 2005
© E.S.I.F.T. srl - Firenze ISSN 1120-009X
INTRODUCTION
The members of the Enterococcus genus are
Gram-positive, catalase-negative cocci and present in
the normal flora of the gastrointestinal tract.
However, they can be significant pathogens causing
endocarditis, urinary tract, bloodstream and wound
infections, especially in elderly patients with serious
underlying diseases and immunocompromised
patients who have been hospitalized for prolonged
periods, use invasive devices or have received broad
spectrum antimicrobial therapy. Recently, an
increase in the occurrence of vancomycin-resistant
enterococci (VRE) has been reported
1,2
. Six types
of glycopeptide resistance have already been
described among enterococci. Three of them are the
most common: VanA, VanB, and VanC pheno-
types. The genes associated with resistance have the
same names as the phenotype, vanA, vanB and
vanC, respectively. The vanA and vanB genes are
found exclusively in Enterococcus faecium and E.
faecalis, which are commonly isolated from human
diseases. VanA-mediated resistance is associated
with inducible high-level resistance to vancomycin
The Resazurin Microplate Method for Rapid Detection
of Vancomycin Resistance in Enterococci
A.Y. COBAN
1
- O. DARKA
1
- N.T. FISGIN
2
- C.C. CIHAN
1
- K. BILGIN
1
- A. AKGUNES
1
T. GUVEN
3
- B. DOKUZOGUZ
3
- A. BIRINCI
1
- B. DURUPINAR
1
1
Ondokuz Mayis University, Medical School, Department of Microbiology and Clinical Microbiology, Samsun, Turkey.
2
Ondokuz Mayis University, Medical School, Department of Clinical Microbiology and Infectious Diseases, Samsun, Turkey.
3
Ankara Numune Education and Research Hospital, The First Infectious Diseases and Clinical Microbiology Clinic, Ankara,
Turkey.
Corresponding author: Ahmet Yilmaz Coban, Ph.D., Ondokuz Mayis University, Medical School,
Department of Microbiology and Clinical Microbiology, 55139 Samsun, Turkey.
Tel: +90 362 4576000/3526; Fax: +90 362 4576041. E-mail: noahmet@yahoo.com
Summary
Vancomycin-resistant enterococci (VRE) are a serious challenge for physicians
because of the limited treatment options for infections caused by this organism.
Prevention of VRE transmission in hospitals requires early detection of infected or
colonized patients. Therefore rapid and correct detection of vancomycin resistance
is essential. In this study, we use the resazurin microplate method (RMM), which is
a modification of the NCCLS and BSAC broth microdilution methods to rapidly
determine the susceptibilities of clinical enterococci isolates to vancomycin. The
alteration in the RMM was relevant to the final bacterial count. In this method,
inoculum that was 10-fold higher than standard methods was used. A total of 80
enterococci, including 11 VRE isolates and 6 vancomycin intermediate isolates,
were screened with this modified colorimetric broth microdilution method. After 4 h
of incubation 30 µl of 0.01% resazurin solution were added to each well and the
plates were reincubated for color change for 5-10 min. The MICs were obtained at
the 4
th
h. The results were in exact agreement with the NCCLS and the BSAC
microdilution methods. Absolute and essential agreements were 100% and there
were no minor, major or very major errors. In conclusion, this modified colorimetric
broth microdilution method can be used as a reliable, easy, cheap and rapid
method for early detection of VRE. Moreover, this method has the potential of
being used to test the susceptibilities of different bacteria to other antibiotics.
Key words: Enterococci, resazurin microplate method, vancomycin, VRE.
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