Evaluation of Rapid Diagnosis of Candidemia by Agglutination Test Using Gold Nanoparticles

Document Type : Original Article (s)

Authors

1 MSc Student of Medical Mycology, Department of Mycology and Parasitology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

2 Professor, Department of Mycology and Parasitology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

3 PhD in Medical Parasitology, Shahrekord Blood Transfusion Organization, Shahrekord, Shahrekord, Iran

4 MSc student of Medical Mycology, Mycology and Parasitology Department, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

5 Associate Professor, Department of Mycology and Parasitology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

Abstract

Background: Candidemia showed high morbidity and mortality and should be diagnosed immediately. The aim of this study was to provide a simple and rapid method for direct detection of Candida in the blood, using conjugated antibodies with gold nanoparticles by the agglutination method.
Methods: In this study, yeast species isolated from 40 blood samples of patients tested by the BACTEC method were used.  After identifying the etiologic agents by morphological and molecular methods, a mixture of yeast antigens was prepared. Briefly, a mixture of antigens was prepared from the Candida isolates detected from candidemia, and injected into healthy rabbit skin with complete adjuvant to produce antibodies. After injecting the incomplete adjuvant in four reminders, blood samples and serum were finally prepared from the rabbit. Antibody production and its efficacy were confirmed by ELISA method. Gold nanoparticles were chemically conjugated with antibodies. For rapid diagnosis of candidemia, the nanoparticle method was examined on the blood of four patients and blood contaminated with a certain number of Candida yeasts in the laboratory.
Findings: Candida albicans was the most common etiologic agent of candidemia, followed by Candida glabrata and Candida parapsilosis complex. In determining the cut-off nanoparticle method, the least yeast detectable in the patient's blood was one yeast. The sensitivity and specificity of the nanoparticle method for detecting Candida in the blood of four patients compared to blood culture was 100%.
Conclusion: Under our laboratory conditions, if testing 15 µl blood of a suspected patient to candidemia containing one yeast or even just Candida antigens exposed to 35 microliters of the nanoparticles conjugated antibodies, it shows agglutination during one minute.

Keywords

References

  1. Calandra T, Roberts JA, Antonelli M, Bassetti M, Vincent JL. Diagnosis and management of invasive candidiasis in the ICU: an updated approach to an old enemy. Crit Care 2016; 20(1): 125.
  2. He S, Hang JP, Zhang L, Wang F, Zhang DC, Gong FH. A systematic review and meta-analysis of diagnostic accuracy of serum 1, 3-β-D-glucan for invasive fungal infection: focus on cutoff levels.
    J Microbiol Immunol Infect 2015; 48(4): 351-61.
  3. Lamoth F, Cruciani M, Mengoli C, Castagnola E, Lortholary O, Richardson M, et al. β-Glucan antigenemia assay for the diagnosis of invasive fungal infections in patients with hematological malignancies: a systematic review and meta-analysis of cohort studies from the Third European Conference on Infections in Leukemia (ECIL-3). Clin Infect Dis 2012; 54(5): 633-43.
  4. Willinger B. Laboratory diagnosis and therapy of invasive fungal infections. Curr Drug Targets 2006; 7(4): 513-22.
  5. Mieszawska AJ, Mulder WJM, Fayad ZA, Cormode DP. Multifunctional gold nanoparticles for diagnosis and therapy of disease. Mol Pharm 2013; 10(3): 831-47.
  6. Kumar A, Mazinder Boruah B, Liang XJ. Gold nanoparticles: promising nanomaterials for the diagnosis of cancer and HIV/AIDS. J Nanomater 2011; 2011.
  7. Raghvendra R, Kanthadeivi A, Sathesh K, Aarrthy M. Diagnostics and therapeutic application of gold nanoparticles. Medicine 2014; 6(2): 74-87.
  8. Kianipour S, Emami Ardestani M, Dehghan P. Identification of Candida albicans and Candida dubliniensis species Isolated from bronchoalveolar lavage samples using genotypic and phenotypic methods. Adv Biomed Res 2018; 7: 66.
  9. 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.
  10. Hashemi H, Varshosaz J, Fazeli H, Sharafi SM,
    Mirhendi H, Chadeganipour M, et al. Rapid differential diagnosis of vaginal infections using gold nanoparticles coated with specific antibodies. Med Microbiol Immunol 2019; 208(6): 773-80.
  11. Vaezi A, Fakhim H, Khodavaisy S, Alizadeh A, Nazeri M, Soleimani A, et al. Epidemiological and mycological characteristics of candidemia in Iran: a systematic review and meta-analysis. J Mycol Med 2017; 27(2): 146-52.
  12. Agnelli C, Valerio M, Bouza E, Guinea J, Sukiennik T, Guimarães T, et al. Prognostic factors of Candida spp. bloodstream infection in adults: A nine-year retrospective cohort study across tertiary hospitals in Brazil and Spain. Lancet Reg Health Am 2022; 6: 100117.
  13. Charsizadeh A, Mirhendi H, Nikmanesh B, Eshaghi H, Rahmani M, Farhang A, et al. Candidemia in children caused by uncommon species of Candida. Arch Pediatr Infect Dis 2018; 6(2): e11895.
  14. Jabari SK, Chadeganipour M, Ghahri M, Mohammadi R. Etiologic agents of Candidemia in pediatric immunocompromised patients. Iran J Ped Hematol Oncol 2016; 6(4): 209-15.
  15. Razzaghi R, Momen-Heravi M, Erami M, Nazeri M. Candidemia in patients with prolonged fever in Kashan, Iran. Curr Med Mycol 2016; 2(3): 20-6.
  16. Alp S, Arikan-Akdagli S, Gulmez D, Ascioglu S, Uzun O, Akova M. Epidemiology of candidaemia in a tertiary care university hospital: 10‐year experience with 381 candidaemia episodes between 2001 and 2010. Mycoses 2015; 58(8): 498-505.
  17. Montagna MT, Caggiano G, Lovero G, De Giglio O, Coretti C, Cuna T, et al. Epidemiology of invasive fungal infections in the intensive care unit: results of a multicenter Italian survey (AURORA Project). Infection 2013; 41(3): 645-53.
  18. Cuenca-Estrella M, Bernal-Martinez L, Buitrago MJ, Castelli MV, Gomez-Lopez A, Zaragoza O, et al. Update on the epidemiology and diagnosis of invasive fungal infection. Int J Antimicrob Agents 2008; 32(Suppl 2): S143-7.
  19. Berenguer J, Buck M, Witebsky F, Stock F, Pizzo PA, Walsh TJ. Lysis-centrifugation blood cultures in the detection of tissue-proven invasive candidiasis disseminated versus single-organ infection. Diagn Microbiol Infect Dis 1993; 17(2): 103-9.
  20. Ness MJ, Vaughan WP, Woods GL. Candida antigen latex test for detection of invasive candidiasis in immunocompromised patients. J Infect Dis 1989; 159(3): 495-502.
  21. Van Burik JH, Leisenring W, Myerson D, Hackman RC, Shulman HM, Sale GE, et al. The effect of prophylactic fluconazole on the clinical spectrum of fungal diseases in bone marrow transplant recipients with special attention to hepatic candidiasis. An autopsy study of 355 patients. Medicine (Baltimore) 1998; 77(4): 246-54.
  22. Singer C, Kaplan MH, Armstrong D. Bacteremia and fungemia complicating neoplastic disease: a study of 364 cases. Am J Med 1977; 62(5): 731-42.
  23. Kami M, Machida U, Okuzumi K, Matsumura T, Mori Si, Hori A, et al. Effect of fluconazole prophylaxis on fungal blood cultures: an autopsy‐based study involving 720 patients with haematological malignancy. Br J Haematol 2002; 117(1): 40-6.
  24. Thorn JL, Gilchrist KB, Sobonya RE, Gaur NK, Lipke PN, Klotz SA. Postmortem candidaemia: marker of disseminated disease. J Clin Pathol 2010; 63(4): 337-40.
  25. Coletta G, Amendola V. Numerical modelling of the
    optical properties of plasmonic and latex nanoparticles to improve the detection limit of immuno-turbidimetric assays. Nanomaterials (Basel) 2021; 11(5): 1147.
  26. Becker K, Almasri AS, Von Eiff C, Peters G, Heilmann C, Fegeler W. Systematic survey of nonspecific agglutination by Candida spp. in latex assays. J Clin Microbiol 2007; 45(4): 1315-8.
  27. Bansod S, Bonde S, Tiwari V, Bawaskar M, Deshmukh S, Gaikwad S, et al. Bioconjugation of gold and silver nanoparticles synthesized by Fusarium oxysporum and their use in rapid identification of Candida species by using bioconjugate-nano-polymerase chain reaction.
    J Biomed Nanotechnol 2013; 9(12): 1962-71.
  28. Einloo A, Dehghan P, Saluti M, Mirzaahmadi S. Specific identification of candida glabrata via colorimetric assay based on gold nanoparticles.
    J Isfahan Dent Sch 2015; 33(325): 231-41. [In Persian].
  29. Zhao F, Niu L, Yan L, Nong J, Wang C, Wang J, et al. Establishment and application of multiple cross displacement amplification coupled with lateral flow biosensor (MCDA-LFB) for visual and rapid detection of Candida albicans in clinical samples. Front Cell Infect Microbiol 2019; 9: 102.
  30. Hu S, Kang H, Gu F, Wang C, Cheng S, Gong W, et al. Rapid detection method for pathogenic Candida captured by magnetic nanoparticles and identified using SERS via AgNPs+. Int J Nanomedicine 2021; 16: 941-50.
Journal of Isfahan Medical School
Volume 40, Issue 668
4th Week, June
May and June 2022
Pages 248-255

Files

History

  • Receive Date: 25 January 2022
  • Revise Date: 03 July 2022
  • Accept Date: 29 May 2022

Share

How to cite

Statistics