Identification of Candida Species from Clinical Samples in a Honduran Tertiary Hospital
Abstract
1. Introduction
2. Materials and Methods
2.1. Clinical Samples and Yeast Isolation Conditions
2.2. Phenotypic Identification of Yeast Species
2.3. In Silico Analysis of Restriction Fragments
2.4. DNA Extraction
2.5. PCR-RFLP
2.6. Data Analysis
2.7. Long-Term Preservation of Yeast Cultures
3. Results
3.1. Frequency of Candida Species According to HardyCHROM®
3.2. Comparison of MicroScan®, HardyCHROM®, and PCR-RFLP
3.3. Candida Species Detection and Identification Directly from Clinical Samples
4. Discussion
5. Conclusions
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
- Frias-De-Leon, M.G.; Martinez-Herrera, E.; Acosta-Altamirano, G.; Arenas, R.; Rodriguez-Cerdeira, C. Superficial candidosis by Candida duobushaemulonii: An emerging microorganism. Infect. Genet. Evol. 2019, 75, 103960. [Google Scholar] [CrossRef]
- Santolaya, M.E.; Thompson, L.; Benadof, D.; Tapia, C.; Legarraga, P.; Cortes, C.; Rabello, M.; Valenzuela, R.; Rojas, P.; Rabagliati, R.; et al. A prospective, multi-center study of Candida bloodstream infections in Chile. PLoS ONE 2019, 14, e0212924. [Google Scholar] [CrossRef]
- Yapar, N. Epidemiology and risk factors for invasive candidiasis. Ther. Clin. Risk Manag. 2014, 10, 95–105. [Google Scholar] [CrossRef] [PubMed]
- Durga, C.S.; Gupta, N.; Soneja, M.; Bhatt, M.; Xess, I.; Jorwal, P.; Singh, G.; Ray, A.; Nischal, N.; Ranjan, P.; et al. Invasive fungal infections in critically ill patients: A prospective study from a tertiary care hospital in India. Drug. Discov. Ther. 2018, 12, 363–367. [Google Scholar] [CrossRef] [PubMed]
- Montagna, M.T.; Lovero, G.; Borghi, E.; Amato, G.; Andreoni, S.; Campion, L.; Lo Cascio, G.; Lombardi, G.; Luzzaro, F.; Manso, E.; et al. Candidemia in intensive care unit: A nationwide prospective observational survey (GISIA-3 study) and review of the European literature from 2000 through 2013. Eur. Rev. Med. Pharmacol. Sci. 2014, 18, 661–674. [Google Scholar] [PubMed]
- Hashemi Fesharaki, S.; Aghili, S.R.; Shokohi, T.; Boroumand, M.A. Catheter-related candidemia and identification of causative Candida species in patients with cardiovascular disorder. Curr. Med. Mycol. 2018, 4, 7–13. [Google Scholar] [CrossRef]
- Sims, C.R.; Ostrosky-Zeichner, L.; Rex, J.H. Invasive candidiasis in immunocompromised hospitalized patients. Arch. Med. Res. 2005, 36, 660–671. [Google Scholar] [CrossRef]
- Suleyman, G.; Alangaden, G.J. Nosocomial Fungal Infections: Epidemiology, Infection Control, and Prevention. Infect. Dis. Clin. N. Am. 2016, 30, 1023–1052. [Google Scholar] [CrossRef]
- Marol, S.; Yucesoy, M. Molecular epidemiology of Candida species isolated from clinical specimens of intensive care unit patients. Mycoses 2008, 51, 40–49. [Google Scholar] [CrossRef]
- Pfaller, M.A.; Diekema, D.J. Epidemiology of invasive mycoses in North America. Crit. Rev. Microbiol. 2010, 36, 1–53. [Google Scholar] [CrossRef]
- Tsai, M.H.; Hsu, J.F.; Yang, L.Y.; Pan, Y.B.; Lai, M.Y.; Chu, S.M.; Huang, H.R.; Chiang, M.C.; Fu, R.H.; Lu, J.J. Candidemia due to uncommon Candida species in children: New threat and impacts on outcomes. Sci. Rep. 2018, 8, 15239. [Google Scholar] [CrossRef] [PubMed]
- Bhattacharjee, P. Epidemiology and antifungal susceptibility of Candida species in a tertiary care hospital, Kolkata, India. Curr. Med. Mycol. 2016, 2, 20–27. [Google Scholar] [CrossRef] [PubMed]
- Lewis, R.E. Overview of the changing epidemiology of candidemia. Curr. Med. Res. Opin. 2009, 25, 1732–1740. [Google Scholar] [CrossRef] [PubMed]
- Huttunen, R.; Attman, E.; Aittoniemi, J.; Outinen, T.; Syrjanen, J.; Karki, T.; Lyytikainen, O. Nosocomial bloodstream infections in a Finnish tertiary care hospital: A retrospective cohort study of 2175 episodes during the years 1999–2001 and 2005–2010. Infect. Dis. 2015, 47, 20–26. [Google Scholar] [CrossRef] [PubMed]
- Lass-Florl, C. The changing face of epidemiology of invasive fungal disease in Europe. Mycoses 2009, 52, 197–205. [Google Scholar] [CrossRef]
- Clancy, C.J.; Nguyen, M.H. Finding the ‘‘missing 50%’’ of invasive candidiasis: How nonculture diagnostics will improve understanding of disease spectrum and transform patient care. Clin. Infect. Dis. 2013, 56, 1284–1292. [Google Scholar] [CrossRef]
- Vatanshenassan, M.; Boekhout, T.; Lass-Florl, C.; Lackner, M.; Schubert, S.; Kostrzewa, M.; Sparbier, K. Proof of Concept for MBT ASTRA, a Rapid Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry (MALDI-TOF MS)-Based Method To Detect Caspofungin Resistance in Candida albicans and Candida glabrata. J. Clin. Microbiol. 2018, 56, e00420–e00518. [Google Scholar] [CrossRef]
- Bal, A.M.; McGill, M. Rapid species identification of Candida directly from blood culture broths by Sepsityper-MALDI-TOF mass spectrometry: Impact on antifungal therapy. J. R. Coll. Physicians Edinb. 2018, 48, 114–119. [Google Scholar] [CrossRef]
- 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, 21–28. [Google Scholar] [CrossRef]
- Sanglard, D. Emerging Threats in Antifungal-Resistant Fungal Pathogens. Front. Med. (Lausanne) 2016, 3, 11. [Google Scholar] [CrossRef]
- Ortiz, B.; Perez-Aleman, E.; Galo, C.; Fontecha, G. Molecular identification of Candida species from urinary infections in Honduras. Rev. Iberoam. Micol. 2018, 35, 73–77. [Google Scholar] [CrossRef] [PubMed]
- Nucci, M.; Queiroz-Telles, F.; Alvarado-Matute, T.; Tiraboschi, I.N.; Cortes, J.; Zurita, J.; Guzman-Blanco, M.; Santolaya, M.E.; Thompson, L.; Sifuentes-Osornio, J.; et al. Epidemiology of candidemia in Latin America: A laboratory-based survey. PLoS ONE 2013, 8, e59373. [Google Scholar] [CrossRef] [PubMed]
- Mirhendi, H.; Makimura, K.; Khoramizadeh, M.; Yamaguchi, H. A one-enzyme PCR-RFLP assay for identification of six medically important Candida species. Nihon Ishinkin Gakkai Zasshi 2006, 47, 225–229. [Google Scholar] [CrossRef] [PubMed]
- Fazeli, A.; Kordbacheh, P.; Nazari, A.; Daie Ghazvini, R.; Mirhendi, H.; Safara, M.; Bakhshi, H.; Yaghoubi, R. Candiduria in Hospitalized Patients and Identification of Isolated Candida Species by Morphological and Molecular Methods in Ilam, Iran. Iran J. Public Health 2019, 48, 156–161. [Google Scholar] [PubMed]
- Erazo, B.M.; Ramirez, G.A.; Cerrato, L.E.; Pinto, L.J.; Castro, E.J.; Yanez, N.J.; Montoya, B.; Fontecha, G.A. Prevalence of Hb S (HHB: C.20A > T) in a Honduran population of African descent. Hemoglobin 2015, 39, 134–137. [Google Scholar] [CrossRef]
- Jafari, Z.; Motamedi, M.; Jalalizand, N.; Shokoohi, G.R.; Charsizadeh, A.; Mirhendi, H. Comparison of CHROMagar, polymerase chain reaction-restriction fragment length polymorphism, and polymerase chain reaction-fragment size for the identification of Candida species. Curr. Med. Mycol. 2017, 3, 10–15. [Google Scholar] [CrossRef]
- Rezazadeh, E.; Moazeni, M.; Sabokbar, A. Use of cost effective and rapid molecular tools for identification of Candida species, opportunistic pathogens. Curr. Med. Mycol. 2016, 2, 1–4. [Google Scholar] [CrossRef][Green Version]
- Camacho-Cardoso, J.L.; Martinez-Rivera, M.A.; Manzano-Gayosso, P.; Mendez-Tovar, L.J.; Lopez-Martinez, R.; Hernandez-Hernandez, F. Molecular detection of candida species from hospitalized patient’s specimens. Gac. Med. Mex. 2017, 153, 581–589. [Google Scholar] [CrossRef]
- Kaur, R.; Dhakad, M.S.; Goyal, R.; Haque, A.; Mukhopadhyay, G. Identification and Antifungal Susceptibility Testing of Candida Species: A Comparison of Vitek-2 System with Conventional and Molecular Methods. J. Glob. Infect. Dis. 2016, 8, 139–146. [Google Scholar] [CrossRef]
- Dagi, H.T.; Findik, D.; Senkeles, C.; Arslan, U. Identification and antifungal susceptibility of Candida species isolated from bloodstream infections in Konya, Turkey. Ann. Clin. Microbiol. Antimicrob. 2016, 15, 36. [Google Scholar] [CrossRef]
- Sadrossadati, S.Z.; Ghahri, M.; Imani Fooladi, A.A.; Sayyahfar, S.; Beyraghi, S.; Baseri, Z. Phenotypic and genotypic characterization of Candida species isolated from candideamia in Iran. Curr. Med. Mycol. 2018, 4, 14–20. [Google Scholar] [CrossRef] [PubMed]
- Jia, X.; Li, C.; Cao, J.; Wu, X.; Zhang, L. Clinical characteristics and predictors of mortality in patients with candidemia: A six-year retrospective study. Eur. J. Clin. Microbiol. Infect. Dis. 2018, 37, 1717–1724. [Google Scholar] [CrossRef] [PubMed]
- Berrio, I.; Maldonado, N.; De Bedout, C.; Arango, K.; Cano, L.E.; Valencia, Y.; Jimenez-Ortigosa, C.; Perlin, D.S.; Gomez, B.L.; Robledo, C.; et al. Comparative study of Candida spp. isolates: Identification and echinocandin susceptibility in isolates obtained from blood cultures in 15 hospitals in Medellin, Colombia. J. Glob. Antimicrob. Resist. 2018, 13, 254–260. [Google Scholar] [CrossRef] [PubMed]
- Zhai, Y.; Liu, J.; Zhou, L.; Ji, T.; Meng, L.; Gao, Y.; Liu, R.; Wang, X.; Li, L.; Lu, B.; et al. Detection of Candida species in pregnant Chinese women with a molecular beacon method. J. Med. Microbiol. 2018, 67, 783. [Google Scholar] [CrossRef] [PubMed]
- Zarrinfar, H.; Kaboli, S.; Dolatabadi, S.; Mohammadi, R. Rapid detection of Candida species in bronchoalveolar lavage fluid from patients with pulmonary symptoms. Braz. J. Microbiol. 2016, 47, 172–176. [Google Scholar] [CrossRef]
- Mohammadi, R.; Abdi, S. Molecular identification of Candida species isolated from gastro-oesophageal candidiasis in Tehran, Iran. Gastroenterol. Hepatol. Bed Bench 2015, 8, 288–293. [Google Scholar]
- Ostrosky-Zeichner, L.; Pappas, P.G. Invasive candidiasis in the intensive care unit. Crit. Care Med. 2006, 34, 857–863. [Google Scholar] [CrossRef]
- van Asbeck, E.C.; Clemons, K.V.; Stevens, D.A. Candida parapsilosis: A review of its epidemiology, pathogenesis, clinical aspects, typing and antimicrobial susceptibility. Crit. Rev. Microbiol. 2009, 35, 283–309. [Google Scholar] [CrossRef]
- Reiss, E.; Lasker, B.A.; Lott, T.J.; Bendel, C.M.; Kaufman, D.A.; Hazen, K.C.; Wade, K.C.; McGowan, K.L.; Lockhart, S.R. Genotyping of Candida parapsilosis from three neonatal intensive care units (NICUs) using a panel of five multilocus microsatellite markers: Broad genetic diversity and a cluster of related strains in one NICU. Infect. Genet. Evol. 2012, 12, 1654–1660. [Google Scholar] [CrossRef]
- Ruiz de Alegria Puig, C.; Aguero-Balbin, J.; Fernandez-Mazarrasa, C.; Martinez-Martinez, L. Evaluation of the Vitek-MS system in the identification of Candida isolates from bloodstream infections. Rev. Iberoam. Micol. 2018, 35, 130–133. [Google Scholar] [CrossRef]
- Fatima, A.; Bashir, G.; Wani, T.; Jan, A.; Kohli, A.; Khan, M.S. Molecular identification of Candida species isolated from cases of neonatal candidemia using polymerase chain reaction-restriction fragment length polymorphism in a tertiary care hospital. Indian J. Pathol. Microbiol. 2017, 60, 61–65. [Google Scholar] [CrossRef] [PubMed]
- Alfouzan, W.; Dhar, R.; Albarrag, A.; Al-Abdely, H. The emerging pathogen Candida auris: A focus on the Middle-Eastern countries. J. Infect. Public Health 2019, 12, 451–459. [Google Scholar] [CrossRef] [PubMed]
- Huyke, J.; Martin, R.; Walther, G.; Weber, M.; Kaerger, K.; Bougnoux, M.E.; Elias, J.; Kurzai, O. Candida albicans bloodstream isolates in a German university hospital are genetically heterogenous and susceptible to commonly used antifungals. Int. J. Med. Microbiol. 2015, 305, 742–747. [Google Scholar] [CrossRef] [PubMed]
- Ao, W.; Klonoski, J.; Berlinghoff, E.; Jensen, J.; Afroz, T.; Munns, D.; Lindsey, W.; Denys, G.; Jenison, R. Rapid Detection and Differentiation of Clinically Relevant Candida Species Simultaneously from Blood Culture by Use of a Novel Signal Amplification Approach. J. Clin. Microbiol. 2018, 56, e00982–e01017. [Google Scholar] [CrossRef]
- Ullmann, A.J.; Cornely, O.A.; Donnelly, J.P.; Akova, M.; Arendrup, M.C.; Arikan-Akdagli, S.; Bassetti, M.; Bille, J.; Calandra, T.; Castagnola, E.; et al. ESCMID* guideline for the diagnosis and management of Candida diseases 2012: Developing European guidelines in clinical microbiology and infectious diseases. Clin. Microbiol. Infect. 2012, 18, 1–8. [Google Scholar] [CrossRef]
- Leite Junior, D.P.; Yamamoto, A.C.; Martins, E.R.; Teixeira, A.F.; Hahn, R.C. Species of Candida isolated from anatomically distinct sites in military personnel in Cuiaba, Mato Grosso, Brazil. Bras Dermatol 2011, 86, 675–680. [Google Scholar] [CrossRef]
Candida Species | Length of the ITS1-ITS2 Amplicon (bp) | Restriction Fragment Sizes (bp) |
---|---|---|
C. albicans complex | 538 | 299, 239 |
[Candida] 1 glabrata complex | 880 | 563, 317 |
C. parapsilosis complex | 520 | 520 |
C. tropicalis | 528 | 342, 186 |
C. krusei (Pichia kudriavzevii) | 510 | 262, 248 |
C. kefyr (Kluyveromyces marxianus) | 721 | 721 |
C. guillermondii (Meyerozyma guilliermondii) | 607 | 372, 157, 82 |
[Candida] haemulonii complex | 400 | 400 |
C. catenulata (Diutina catenulata) | 402 | 402 |
C. famata (Debaryomyces hansenii) | 639 | 639 |
[Candida] zeylanoides | 626 | 626 |
[Candida] inconspicua | 455 | 245, 210 |
Clinical Sample | N° of samples (%) | C. albicans Complex | C. glabrata Complex | C. parapsilosis Complex | C. tropicalis | C. krusei | C. kefyr | C. haemulonii Complex | C. guillermondii | Unidentified | Total n° of Yeasts (%) |
---|---|---|---|---|---|---|---|---|---|---|---|
Urine | 63 (37.70) | 25 | 16 | 1 | 20 | 1 | 2 | 65 1 | |||
Sputum | 45 (26.95) | 28 | 6 | 3 | 10 | 1 | 2 | 1 | 51 1 | ||
Vaginal swab | 18 (10.78) | 11 | 5 | 1 | 1 | 1 | 1 | 20 1 | |||
Blood | 12 (7.18) | 2 | 5 | 1 | 2 | 2 | 12 | ||||
Catheter | 9 (5.39) | 3 | 2 | 3 | 1 | 9 | |||||
Stool | 5 (2.99) | 2 | 1 | 2 | 5 | ||||||
Cutaneous secretion | 2 (1.20) | 1 | 1 | 2 | |||||||
Otic secretion | 2 (1.20) | 1 | 1 | 2 | |||||||
Oral swab | 4 (2.39) | 3 | 1 | 4 | |||||||
CSF | 2 (1.20) | 2 | 2 | ||||||||
Abscess | 5 (2.99) | 1 | 1 | 1 | 1 | 1 | 5 | ||||
Total (%) | 167 (100) | 76 (42.93%) | 30 (16.94%) | 16 (9.03%) | 37 (20.9%) | 8 (4.51%) | 5 (2.82%) | 2 (1.12%) | 2 (1.12%) | 1 (0.56%) | 177 (100%) |
Candida species | MicroScan® | HardyCHROM® | PCR-RFLP |
---|---|---|---|
C. albicans complex | 69 | 74 | 76 |
[Candida] glabrata complex | 25 | 29 | 29 |
C. parapsilosis complex | 11 | 10 | 16 |
C. tropicalis | 26 | 32 | 37 |
C. krusei | 10 | 4 | 8 |
C. guillermondii | 11 | 2 | |
[Candida] haemulonii complex | 2 | ||
Candida kefyr | 4 | 5 | |
C. famata | 6 | ||
C. catenulata | 4 | ||
[Candida] inconspicua | 1 | ||
[Candida] zeylanoides | 1 | ||
Total | 168 | 149 | 175 |
Method | PCR-RFLP | HardyCHROM® | MicroScan® |
---|---|---|---|
MicroScan® | 0.648 (0.041; 0.568–0.727) 1 | ||
PCR-RFLP | 0.653 (0.042; 0.572–0.735) 1 | ||
HardyCHROM® | 0.662 (0.039; 0.568–0.739) 1 |
Clinical Sample | N of Clinical Samples (%) | C. albicans Complex | C. glabrata Complex | C. parapsilosis Complex | C. tropicalis | C. krusei | C. haemulonii Complex |
---|---|---|---|---|---|---|---|
Sputum | 13 (28.26) | 8 | 1 | 1 | 3 | ||
Vaginal swab | 12 (26.09) | 5 | 3 | 2 | 1 | 1 | |
Blood | 8 (17.39) | 2 | 4 | 2 | |||
Cutaneous secretion | 2 (4.35) | 1 | 1 | ||||
Otic secretion | 2 (4.35) | 1 | 1 | ||||
Oral swab | 5 (10.87) | 2 | 1 | 1 | 1 | ||
CSF | 2 (4.35) | 2 | |||||
Rectal swab | 2 (4.35) | 2 | |||||
Total | 46 (100) | 19 (41.3) | 5 (10.87) | 10 (21.74) | 8 (17.39) | 2 (4.35) | 2 (4.35) |
Clinical Samples | Axenic Culture | n |
---|---|---|
C. tropicalis | C. albicans complex | 2 |
C. parapsilosis complex | C. albicans complex | 2 |
C. albicans complex | C. tropicalis | 3 |
C. albicans complex | Candida spp. | 1 |
C. tropicalis | C. guillermondii | 1 |
C. albicans complex | C. krusei | 1 |
C. haemulonii complex | C. kefyr | 1 |
C. glabrata complex | C. haemulonii complex | 1 |
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Montes, K.; Ortiz, B.; Galindo, C.; Figueroa, I.; Braham, S.; Fontecha, G. Identification of Candida Species from Clinical Samples in a Honduran Tertiary Hospital. Pathogens 2019, 8, 237. https://doi.org/10.3390/pathogens8040237
Montes K, Ortiz B, Galindo C, Figueroa I, Braham S, Fontecha G. Identification of Candida Species from Clinical Samples in a Honduran Tertiary Hospital. Pathogens. 2019; 8(4):237. https://doi.org/10.3390/pathogens8040237
Chicago/Turabian StyleMontes, Kathy, Bryan Ortiz, Celeste Galindo, Isis Figueroa, Sharleen Braham, and Gustavo Fontecha. 2019. "Identification of Candida Species from Clinical Samples in a Honduran Tertiary Hospital" Pathogens 8, no. 4: 237. https://doi.org/10.3390/pathogens8040237
APA StyleMontes, K., Ortiz, B., Galindo, C., Figueroa, I., Braham, S., & Fontecha, G. (2019). Identification of Candida Species from Clinical Samples in a Honduran Tertiary Hospital. Pathogens, 8(4), 237. https://doi.org/10.3390/pathogens8040237