Identification of Candida species by restriction enzyme analysis
Background/aim: The identification of Candida species isolated from clinical specimens provides information about antifungal
susceptibility and sheds light on the choice of empirical treatment. In the present study, restriction enzyme analysis of C. albicans and
non-albicans Candida species previously identified by conventional methods was done to evaluate the utility of restriction enzyme
analysis for more rapid and reliable identification of Candida species.
Materials and methods: A total of 146 Candida strains isolated from various clinical specimens and ATCC strains were included. PCR
products were digested with MwoI for all species and with BslI for C. parapsilosis and C. tropicalis strains.
Results: The strains were identified by conventional methods as 40 C. albicans, 27 C. parapsilosis, 26 C. tropicalis, 25 C. glabrata, 11 C.
kefyr, 10 C. krusei, and 7 C. guilliermondii strains. Restriction digestion with MwoI was able to distinguish between five different species
(C. albicans, C. krusei, C. guilliermondii, C. kefyr, and C. glabrata), while BslI digestion could distinguish between C. tropicalis and C.
parapsilosis.
Conclusion: Restriction enzyme analysis with MwoI and BslI can be used for the identification of Candida species in situations where
rapid identification is necessary or conventional methods are problematic.
___
- Martin GS, Mannino DM, Eaton S, Moss M. The epidemiology
of sepsis in the United States from 1979 through 2000. N Engl
J Med 2003; 348: 1546-1554.
- Quindós G. Epidemiology of candidaemia and invasive
candidiasis. A changing face. Rev Iberoam Micol 2014; 31: 42-
48.
- Kullberg BJ, Arendrup MC. Invasive candidiasis. N Engl J Med
2015; 373: 1445-1456
- Gudlaugsson O, Gillespie S, Lee K, Vande Berg J, Hu J, Messer
S, Herwaldt L, Pfaller M, Diekema D. Attributable mortality
of nosocomial candidemia, revisited. Clin Infect Dis 2003; 37:
1172-1177.
- Marchetti O, Bille J, Fluckiger U, Eggimann P, Ruef C,
Garbino J, Calandra T, Glauser MP, Täuber MG, Pittet D et al.
Epidemiology of candidemia in Swiss tertiary care hospitals:
secular trends, 1991–2000. Clin Infect Dis 2004; 38: 311-320.
- Vallabhaneni S, Mody RK, Walker T, Chiller T. The global
burden of fungal diseases. Infect Dis Clin N Am 2016; 30: 1-11.
- Concia E, Azzini AM, Conti M. Epidemiology, incidence
and risk factors for invasive candidiasis in high-risk patients.
Drugs 2009; 69: 5-14.
- J. Guinea. Global trends in the distribution of
Candida
species
causing candidemia. Clin Microbiol Infect 2014; 20: 5-10.
- Orasch C, Marchetti O, Garbino J, Schrenzel J, Zimmerli
S, Muhlethaler K, Pfyffer G, Ruef C, Fehr J, Zbinden R et al.
Candida
species distribution and antifungal susceptibility
testing according to European Committee on Antimicrobial
Susceptibility Testing and new vs. old Clinical and Laboratory
Standards Institute clinical breakpoints: a 6-year prospective
candidaemia survey from the fungal infection network of
Switzerland. Clin Microbiol Infect 2014; 20: 698-705
- Sardi JC, Scorzoni L, Bernardi T, Fusco-Almeida AM, Mendes
Giannini MJ.
Candida
species: current epidemiology,
pathogenicity, biofilm formation, natural antifungal products
and new therapeutic options. J Med Microbiol 2013; 62: 10-24.
- Peman J, Bosch M, Canton E, Viudes A, Jarque I, Gómez-
García M, García-Martínez JM, Gobernado M. Fungemia due
to
Candida guilliermondii
in a pediatric and adult population
during a 12-year period. Diagn Microbiol Infect Dis 2008; 60:
109-112.
- Peman J, Zaragoza R. Current diagnostic approaches to
invasive candidiasis in critical care settings. Mycoses 2010; 53:
424-433.
- Trost A, Graf B, Eucker J, Sezer O, Possinger K, Göbel UB,
Adam T. Identification of clinically relevant yeasts by PCR/
RFLP. J Microbiol Meth 2004; 56: 201-211.
- Leaw SN, Chang HC, Sun HF, Barton R, Bouchara JP, Chang
TC. Identification of medically important yeast species by
sequence analysis of the internal transcribed spacer regions. J
Clin Microbiol 2006; 44: 693-699.
- Cassagne C, Cella AL, Suchon P, Normand AC, Ranque
S, Piarroux R. Evaluation of four pretreatment procedures for
MALDI-TOF MS yeast identification in the routine clinical
laboratory. Med Mycol 2013; 51: 371-377.
- Alam MZ, Alam Q, Jiman-Fatani A, Kamal MA, Abuzenadah
AM, Chaudhary AG, Akram M, Haque A.
Candida
identification: a journey from conventional to molecular
methods in medical mycology. World J Microbiol Biotechnol
2014; 30: 1437-1451
- Tekeli A, Akan OA, Koyuncu E, Dolapci I, Uysal S. Initial
Candida dubliniensis
isolate in
Candida
spp. positive
haemocultures in Turkey between 2001 and 2004. Mycoses
2006; 49: 60-64.
- Vijayakumar R, Giri S, Kindo A. Molecular species
identification of
Candida
from blood samples of intensive care
unit patients by polymerase chain reaction: restricted fragment
length polymorphism. J Lab Phys 2012; 4: 1.
- Khodadadi H, Karimi L, Jalalizand N, Adin H, Mirhendi H.
Utilization of size polymorphism in ITS1 and ITS2 regions
for identification of pathogenic yeast species. J Med Microbiol
2017; 66: 126-133.
- Williams DW, Wilson MJ, Lewis MA, Potts AJ. Identification
of
Candida
species by PCR and restriction fragment length
polymorphism analysis of intergenic spacer regions of
ribosomal DNA. J Clin Microbiol 1995; 33: 2476-2479.
- Shokohi T, Soteh MB, Saltanat Pouri Z, Hedayati MT, Mayahi
S. Identification of
Candida
species using PCR-RFLP in cancer
patients in Iran. Indian J Med Microbiol 2010; 28: 147-151.
- Mirhendi H, Makimura K, Khoramizadeh M, Yamaguchi
H. A one-enzyme PCR-RFLP assay for identification of six
medically important
Candida
species. Jpn J Med Mycol 2006;
47: 225-229.
- Irobi J, Schoofs A, Goossens H. Genetic identification of
Candida
species in HIV-positive patients using the polymerase
chain restriction fragment length polymorphism analysis of its
DNA. Mol Cell Probes 1999; 13: 401-406.
- Pinto PM, Resende MA, Koga-Ito CY, Ferreira JA, Tendler
M. rDNA-RFLP identification of
Candida
species in
immunocompromised and seriously diseased patients. Can J
Microbiol 2004; 50: 504-520.
- Fanci R, Pecile P. Central venous catheter-related infection
due to
Candida membranaefaciens
, a new opportunistic azole-
resistant yeast in a cancer patient: a case report and a review of
literature. Mycoses 2005; 48: 357-359.
- Aghili SR, Shokohi T, Boroumand MA, Hashemi Fesharaki
S, Salmanian B. Intravenous catheter-associated candidemia
due to
Candida membranaefaciens
: the first Iranian case. J
Tehran Heart Cent 2015; 10: 101-105.