Identification of invasive fungal infections in patients with hematologic disorders by Real-time PCR

Document Type : Research Paper

Authors

1 Associate Professor, Department of Parasitology and Mycology, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

2 Master of Sciences, Department of Parasitology and Mycology, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

3 Associate Professor, Department of Pediatrics, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

4 Associate Professor, Department of Social Medicine, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

5 , Department of Pediatrics, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

6 Professor, Alborzi Clinical Microbiology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran

Abstract

Introduction: Invasive fungal infections due to Aspergillus and Candida often occur in leukemia and transplantation patients because of a severe compromised in their immune systems. Rapid identification and initiation of treatment in the early stages of the disease can reduce mortality from fungal infection. The aim of this study was the identification of invasive fungal infections by Real-time PCR in patients with hematologic disorders and bone marrow transplantation.
Methods and Materials: patients with immunodeficiency hospitalized in Mashhad hospitals between March to November 2018 were evaluated for invasive fungal infections using TaqMan Real-time PCR with fluorescence probes and specific primers to identify fungal DNA in peripheral blood samples.
Results: Seventy-five patients participated in this study. The female/ male ratio was 34 (45.3%)/ 41 (54.7%). The most common underlying disease was acute lymphoblastic leukemia (45.3%) and acute myeloid leukemia (32%) and the lowest was related to haemophagocytic patients with lymphohistiocytosis (1.3%). Blood culture results showed isolation of non-albicans Candida (13.3%) and Candida albicans (1.3%). Aspergillus species was not isolated from the blood culture. Real-time PCR were positive for Candida albicans and Aspergillus species in 5 (6.7%) and 9 patients (12%), respectively and 61 patients (81.3%) had negative results for invasive fungal infections.
Conclusion: Fungi are one of the major cause of infections in the patients with hematologic disorders. Real-time PCR techniques can be a reliable and fast way to diagnose these infections and manage the disease in high-risk individuals.

Keywords


  1. Pfaller MA, Diekema DJ. Epidemiology of invasive mycoses in North America. Critical reviews in
    microbiology. 2010;36(1):1-53.
    2. Horn DL, Neofytos D, Anaissie EJ, Fishman JA, Steinbach WJ, Olyaei AJ, et al. Epidemiology
    and outcomes of candidemia in 2019 patients: data from the prospective antifungal therapy alliance
    registry. Clinical infectious diseases. 2009;48(12):1695-703.
    3. Badiee P, Kordbacheh P, Alborzi A, Malekhoseini SA. Invasive fungal infection in renal
    transplant recipients demonstrated by panfungal polymerase chain reaction. Exp Clin Transplant.
    2007;5(1):624-9.
    4. Blumberg HM, Jarvis WR, Soucie JM, Edwards JE, Patterson JE, Pfaller MA, et al. Risk factors
    for candidal bloodstream infections in surgical intensive care unit patients: the NEMIS prospective
    multicenter study. Clinical Infectious Diseases. 2001;33(2):177-86.
    5. Wisplinghoff H, Bischoff T, Tallent SM, Seifert H, Wenzel RP, Edmond MB. Nosocomial
    bloodstream infections in US hospitals: analysis of 24,179 cases from a prospective nationwide
    surveillance study. Clinical infectious diseases. 2004;39(3):309-17.
    6. Dornbusch HJ, Manzoni P, Roilides E, Walsh TJ, Groll AH. Invasive fungal infections in
    children.Pediatr Infect Dis J 2009;28(8):734-7.
    7. Bille J, Marchetti O, Calandra T. Changing face of health-care associated fungal infections. Curr
    Opin Infect Dis 2005;18:314-9.
    8. Garcia-Vidal C, Upton A, Kirby KA, Marr KA. Epidemiology of invasive mold infections in
    allogeneic stem cell transplant recipients: biological risk factors for infection according to time after
    transplantation. Clinical Infectious Diseases. 2008;47(8):1041-50.
    9. Neofytos D, Fishman JA, Horn D, Anaissie E, Chang CH, Olyaei A, et al. Epidemiology and
    outcome of invasive fungal infections in solid organ transplant recipients. Transplant Infectious Disease.
    2010;12(3):220-9.
    10. Dimopoulos G, Matthaiou DK, Moussas N, Apostolopoulou O, Arabatzis G, Velegraki A, et al.
    Pathogenesis of Aspergillosis in Humans. Human Emerging and Reā€emerging Infections: Viral and
    Parasitic Infections. 2015:967-84.
    11. Perfect JR, Cox GM, Lee JY, Kauffman CA, De Repentigny L, Chapman SW, et al. The impact
    of culture isolation of Aspergillus species: a hospital-based survey of aspergillosis. Clinical Infectious
    Diseases. 2001;33(11):1824-33.
    12. Schabereiter-Gurtner C, Selitsch B, Rotter ML, Hirschl AM, Willinger B. Development of novel
    real-time PCR assays for detection and differentiation of eleven medically important Aspergillus and
    Candida species in clinical specimens. Journal of clinical microbiology. 2007;45(3):906-14.
    13. Badiee P, Alborzi A, Shakiba E, Ziyaeyan M, Pourabbas B. Molecular diagnosis of Aspergillus
    endocarditis after cardiac surgery. Journal of medical microbiology. 2009;58(2):192-5.
    14. Bretagne S, Costa JM. Towards a molecular diagnosis of invasive aspergillosis and disseminated
    candidosis. FEMS Immunol Med Microbiol 2005;45(3):361-8.
    15. De Pauw B, Walsh TJ, Donnelly JP, Stevens DA, Edwards JE, Calandra T, et al. Revised
    definitions of invasive fungal disease from the European organization for research and treatment of
    cancer/invasive fungal infections cooperative group and the national institute of allergy and infectious
    diseases mycoses study group (EORTC/MSG) consensus group. Clinical infectious diseases.
    2008;46(12):1813-21.
    16. Choi J-K, Cho S-Y, Yoon S-S, Moon J-H, Kim S-H, Lee J-H, et al. Epidemiology and Risk
    Factors for Invasive Fungal Diseases among Allogeneic Hematopoietic Stem Cell Transplant Recipients
    in Korea: Results of “RISK” Study. Biology of Blood and Marrow Transplantation. 2017;23(10):1773-9.
    17. Yapar N. Epidemiology and risk factors for invasive candidiasis. Therapeutics and clinical risk
    management. 2014;10:95.
    18. Badiee P, Badali H, Boekhout T, Diba K, Moghadam AG, Nasab AH, et al. Antifungal
    susceptibility testing of Candida species isolated from the immunocompromised patients admitted to ten
    university hospitals in Iran: comparison of colonizing and infecting isolates. BMC infectious diseases.
    2017;17(1):727.
    19. Mello E, Posteraro B, Vella A, De Carolis E, Torelli R, D'Inzeo T, et al. Susceptibility testing of
    common and uncommon Aspergillus species against posaconazole and other mold-active antifungal
    azoles using the Sensititre method. Antimicrobial agents and chemotherapy. 2017:AAC. 00168-17.
  2. 20. Badiee P, Alborzi A, Moeini M, Haddadi P. Antifungal susceptibility of the Aspergillus species
    by Etest and CLSI reference methods. Archives of Iranian medicine. 2012;15(7):429.
    21. Alam FF, Mustafa AS, Khan ZU. Comparative evaluation of (1, 3)-β-D-glucan, mannan and
    anti-mannan antibodies, and Candida species-specific snPCR in patients with candidemia. BMC
    infectious diseases. 2007;7(1):103.
    22. Hao W, Pan Y-X, Ding Y-Q, Xiao S, Yin K, Wang Y-D, et al. Well-characterized monoclonal
    antibodies against cell wall antigen of Aspergillus species improve immunoassay specificity and
    sensitivity. Clinical and Vaccine Immunology. 2008;15(2):194-202.
    23. Mikulska M, Furfaro E, Viscoli C. Biomarkers for diagnosis and follow-up of invasive
    candidiasis: a brief review of the ECIL recommendations. Current Fungal Infection Reports.
    2012;6(3):192-7.
    24. White PL, Archer AE, Barnes RA. Comparison of non-culture-based methods for detection of
    systemic fungal infections, with an emphasis on invasive Candida infections. Journal of clinical
    microbiology. 2005;43(5):2181-7.
    25. White PL, Perry MD, Barnes RA. An update on the molecular diagnosis of invasive fungal
    disease. FEMS microbiology letters. 2009;296(1):1-10.
    26. Badiee P, Kordbacheh P, Alborzi A, Zakernia M, Haddadi P. Early detection of systemic
    candidiasis in the whole blood of patients with hematologic malignancies. Jpn J Infect Dis. 2009;62(1):1-
    5.
    27. Hope W, Morton A, Eisen DP. Increase in prevalence of nosocomial non-Candida albicans
    candidaemia and the association of Candida krusei with fluconazole use. Journal of Hospital Infection.
    2002;50(1):56-65.
    28. Girmenia C, Martino P, De Bernardis F, Gentile G, Boccanera M, Monaco M, et al. Rising
    incidence of Candida parapsilosis fungemia in patients with hematologic malignancies: clinical aspects,
    predisposing factors, and differential pathogenicity of the causative strains. Clinical Infectious Diseases.
    1996;23(3):506-14.
    29. Trofa D, Gácser A, Nosanchuk JD. Candida parapsilosis, an emerging fungal pathogen. Clinical
    microbiology reviews. 2008;21(4):606-25.
    30. Tan TY, Tan AL, Tee N, Ng L. A retrospective analysis of antifungal susceptibilities of Candida
    bloodstream isolates from Singapore hospitals. Ann Acad Med Singapore. 2008;37(10):835-40.
    31. Pagano L, Caira M, Candoni A, Offidani M, Fianchi L, Martino B, et al. The epidemiology of
    fungal infections in patients with hematologic malignancies: the SEIFEM-2004 study. Haematologica.
    2006;91(8):1068-75.
    32. Zaoutis TE, Heydon K, Chu JH, Walsh TJ, Steinbach WJ. Epidemiology, outcomes, and costs of
    invasive aspergillosis in immunocompromised children in the United States, 2000. Pediatrics.
    2006;117(4):e711-e6.
    33. Badiee P, Zareifar S, Haddadi P, Jafarian H. Incidence of Fungal Infections in Pediatric
    Patients with Hematologic Neoplasms. Archives of Pediatric Infectous Disease .2017;5(3): :e41317.