Volume 3, Issue 3 (September 2017)                   Curr Med Mycol 2017, 3(3): 10-15 | Back to browse issues page


XML Print


Download citation:
BibTeX | RIS | EndNote | Medlars | ProCite | Reference Manager | RefWorks
Send citation to:

Jafari Z, Motamedi M, Jalalizand N, Shokoohi G R, Charsizadeh A, Mirhendi H. A comparison between CHROMagar, PCR-RFLP and PCR-FSP for identification of Candida species. Curr Med Mycol. 2017; 3 (3) :10-15
URL: http://cmm.mazums.ac.ir/article-1-167-en.html
Abstract:   (745 Views)
Background and Purpose: The epidemiological alteration in the distribution of Candida species, as well as the significantly increasing trend of either intrinsic or acquired resistance of some of these fungi highlights the need for a reliable method for the identification of the species. Polymerase chain reaction (PCR) is one of the methods facilitating the quick and precise identification of Candida species. The aim of this study was to compare the efficiency of CHROMagar, PCR-restriction fragment length polymorphism (PCR-RFLP), and PCR-fragment size polymorphism (PCR-FSP) assays in the identification of Candida species to determine the benefits and limitations of these methods.
Materials and Methods: This study was conducted on 107 Candida strains, including 20 standard strains and 87 clinical isolates. The identification of the isolates was accomplished by using CHROMagar as a conventional method. The PCR-RFLP assay was performed on the entire internal transcribed spacer (ITS) region of ribosomal DNA (rDNA), and the consequent enzymatic digestion was compared with PCR-FSP results in which ITS1 and ITS2 regions were separately PCR amplified. In both molecular assays, yeast identification was carried out through the specific electrophoretic profiles of the PCR products.
Results: According to the results, the utilization of CHROMagar resulted in the identi-fication of 29 (33.3%) Candida isolates, while the PCR-RFLP and PCR-FSP facilitated the identification of 83 (95.4%) and 80 (91.9%) isolates, respectively. The obtained concordances between CHROMagar and PCR-RFLP, between CHROMagar and PCR-FSP, as well as between PCR-RFLP and PCR-FSP were 0.20, 0.23, and 0.77, respectively.
Conclusion: The recognition of the benefits and limitations of PCR methods allows for the selection of the most efficient technique for a fast and correct differentiation. The PCR-RFLP and PCR-FSP assays had satisfactory concordance. The PCR-FSP provides a rapid, technically simple, and cost-effective method for the identification of Candida species. Nevertheless, to accurately differentiate among the taxonomically related species, PCR-RFLP should be implemented.
 
Full-Text [PDF 440 kb]   (338 Downloads)    
Type of Study: Original Articles | Subject: Laboratory Diagnosis
Received: 2017/10/29 | Accepted: 2017/12/25 | Published: 2018/02/17

References
1. Naglik JR, Challacombe SJ, Hube B. Candida albicans secreted aspartyl proteinases in virulence and pathogenesis. Microbiol Mol Biol Rev. 2003; 67(3):400-28. [DOI:10.1128/MMBR.67.3.400-428.2003]
2. Jarvis WR. Epidemiology of nosocomial fungal infections, with emphasis on Candida species. Clin Infect Dis. 1995; 20(6):1526-30. [DOI:10.1093/clinids/20.6.1526]
3. Hajjeh RA, Sofair AN, Harrison LH, Lyon GM, Arthington-Skaggs BA, Mirza SA, et al. Incidence of bloodstream infections due to Candida species and in vitro susceptibilities of isolates collected from 1998 to 2000 in a population-based active surveillance program. J Clin Microbiol. 2004;42(4):1519-27. [DOI:10.1128/JCM.42.4.1519-1527.2004]
4. Ostrosky-Zeichner L, Pappas PG. Invasive candidiasis in the intensive care unit. Crit Care Med. 2006; 34(3):857-63. [DOI:10.1097/01.CCM.0000201897.78123.44]
5. Aikawa NE, Rosa DT, Del Negro GM, Moraes JC, Ribeiro AC, Saad CG, et al. Systemic and localized infection by Candida species in patients with rheumatic diseases receiving anti-TNF therapy. Rev Bras Reumatol. 2015;15:96
6. Chow JK, Golan Y, Ruthazer R, Karchmer AW, Carmeli Y, Lichtenberg D, et al. Factors associated with candidemia caused by non-albicans Candida species versus Candida albicans in the intensive care unit. Clin Infect Dis. 2008; 46(8):1206-13. [DOI:10.1086/529435]
7. Lewis RE. Overview of the changing epidemiology of candidemia. Curr Med Res Opin. 2009; 25(7):1732-40.
8. Beck-Sague C. National Nosocomial Infections Surveillance System. Secular trends in the epidemiology of nosocomial fungal infections in the United States, 1980-1990. J infect Dis. 1993; 167(5):1247-51 [DOI:10.1093/infdis/167.5.1247]
9. Rangel-Frausto MS, Wiblin T, Blumberg HM, Saiman L, Patterson J, Rinaldi M, et al. National epidemiology of mycoses survey (NEMIS): variations in rates of bloodstream infections due to Candida species in seven surgical intensive care units and six neonatal intensive care units. Clin Infect Dis. 1999; 29(2):253-8. [DOI:10.1086/520194]
10. Pfaller M, Diekema D, Jones R, Messer S, Hollis R, Group SP. Trends in antifungal susceptibility of Candida spp. isolated from pediatric and adult patients with bloodstream infections: SENTRY Antimicrobial Surveillance Program, 1997 to 2000. J Clin Microbiol. 2002; 40(3):852-6. [DOI:10.1128/JCM.40.3.852-856.2002]
11. Asmundsdottir LR, Erlendsdottir H, Haraldsson G, Guo H, Xu J, Gottfredsson M. Molecular epidemiology of candidemia: evidence of clusters of smoldering nosocomial infections. Clin Infect Dis. 2008; 47(2):e17-24. [DOI:10.1086/589298]
12. Neu N, Malik M, Lunding A, Whittier S, Alba L, Kubin C, et al. Epidemiology of candidemia at a children's hospital, 2002 to 2006. Pediatr Infect Dis J. 2009; 28(9):806-9. [DOI:10.1097/INF.0b013e3181a0d78d]
13. Liguori G, Di Onofrio V, GallA F, Lucariello A, Albano L, Catania M, et al. Candida albicans identification: comparison among nine phenotypic systems and a multiplex PCR. J Prev Med Hyg. 2010; 51(3):121-2.
14. Elie CM, Lott TJ, Reiss E, Morrison CJ. Rapid identification of Candida species with species-specific DNA probes. J Clin Microbiol. 1998; 36(11):3260-5.
15. Mohammadi R, Badiee P, Badali H, Abastabar M, Safa AH, Hadipour M, et al. Use of restriction fragment length polymorphism to identify Candida species, related to onychomycosis. Advanced biomedical research. 2015; 4:95. [DOI:10.4103/2277-9175.156659]
16. Fathi N, Mohammadi R, Tabatabaiefar MA, Ghahri M, Sadrossadati SZ. Sequence-identification of Candida species isolated from candidemia. Advanced biomedical research. 2016; 5:150. [DOI:10.4103/2277-9175.188485]
17. Zhang J, Hung G-C, Nagamine K, Li B, Tsai S, Lo S-C. Development of Candida-Specific Real-Time PCR Assays for the Detection and Identification of Eight Medically Important Candida Species. Microbiol Insights. 2016; 9:21-8. [DOI:10.4137/MBI.S38517]
18. 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(2):126-33. [DOI:10.1099/jmm.0.000426]
19. Mirhendi H, Makimura K, Khoramizadeh M, Yamaguchi H. A one-enzyme PCR-RFLP assay for identification of six medically important Candida species. Nippon Ishinkin Gakkai Zasshi. 2006; 47(3):225-9. [DOI:10.3314/jjmm.47.225]
20. Shokohi T, Soteh MH, Pouri ZS, Hedayati M, Mayahi S. Identification of Candida species using PCR-RFLP in cancer patients in Iran. Indian J Med Microbiol. 2010; 28(2):147-51 [DOI:10.4103/0255-0857.62493]
21. Mousavi SA, Khalesi E, Bonjar GS, Aghighi S, Sharifi F, Aram F. Rapid molecular diagnosis for-candida species using PCR-RFLP. Biotechnology. 2007; 6(4):583-7. [DOI:10.3923/biotech.2007.583.587]
22. Mirhendi H, Adin H, Shidfar M, Kordbacheh P, Hashemi J, Moazeni M, et al. Identification of pathogenic Candida species: PCR-fragment size polymorphism (PCR-FSP) assay. Tehran Univ Med J. 2008; 66(9):639-45.
23. Mohammadi R, Mirhendi H, Rezaei-Matehkolaei A, Ghahri M, Shidfar MR, Jalalizand N, et al. Molecular identification and distribution profile of Candida species isolated from Iranian patients. Med Mycol. 2013; 51(6):657-63. [DOI:10.3109/13693786.2013.770603]
24. Kazemi A, Falahati M, Hajipoor A, Jafari A. Comparison of Phenotypic Tests and PCR to Detect Candida Albicans From Vaginal Specimens (Tabriz, 2009-2010). Jundishapur J Microbiol. 2013; 6(2):122-6 [DOI:10.5812/jjm.4734]
25. Ahmad S, Khan Z, Mustafa AS, Khan ZU. Seminested PCR for diagnosis of candidemia: comparison with culture, antigen detection, and biochemical assays for species identification. J Clin Microbiol. 2002; 40(7):2483-9. [DOI:10.1128/JCM.40.7.2483-2489.2002]
26. Pfaller MA, Houston A, Coffmann S. Application of CHROMagar Candida for rapid screening of clinical specimens for Candida albicans, Candida tropicalis, Candida krusei, and Candida (Torulopsis) glabrata. J Clin Microbiol. 1996; 34(1):58-61.
27. Souza MN, Ortiz SO, Mello MM, Oliveira FdM, Severo LC, Goebel CS. Comparison between four usual assays of identification of Candida species. Rev Inst Med Trop Sao Paulo. 2015; 57(4):281-7. [DOI:10.1590/S0036-46652015000400002]
28. Estrada-Barraza D, Martinez AD, Flores-Padilla L, Mendoza-De Elias R, Sanchez-Vargas LO. Comparison between conventional methods, ChromAgar Candida® and PCR method for the identification of Candidaspecies in clinical isolates. Rev Iberoam Micol. 2011;28(1):36-42. [DOI:10.1016/j.riam.2010.11.003]

Add your comments about this article : Your username or Email:
CAPTCHA code

Send email to the article author


© 2015 All Rights Reserved | Current Medical Mycology