Articles

Comparison of histopathological analysis, culture and polymerase chain reaction assays to detect mucormycosis in biopsy and blood specimens

Abstract

Background and Objectives: The aim of this study was to compare direct microscopic examination with culture and PCR for the diagnosis of Mucorales infection in blood and tissue specimens.
Material and Methods: Blood samples and tissue specimens were obtained from 28 patients (total 58 samples) with suspected invasive fungal infection and cultured on proper media. Direct smear of tissue samples was done with potassium hydroxide, hematoxylin and eosin, and methenamine silver staining. DNA extracted from blood and tissue specimens were used for semi-nested PCR targeting 18S rDNA of Mucorales species.
Results: Mucormycosis was documented in 7/28 (25%) of tissue specimens with positive findings by direct smear, of which PCR and culture were positive in 6 (86%) and 5 (70%) specimens, respectively. The etiologic agents were Mucor spp. and Rhizopus spp. However, culture and PCR results for all blood specimens were negative.
Conclusions: As the orders of Mucorales do not have well growth in culture media, PCR with tissue specimens is more sensitive than tissue or blood culture methods. Unfortunately, there is no alternative method for direct smear, which is an invasive method. Molecular methods may be helpful in these cases.

Marr KA, Carter RA, Crippa F, Wald A, Corey L.Epidemiology and outcome of mold infections in hematopoietic stem cell transplant recipients. Clin Infect Dis 2002; 34: 909-917.

Sun QN, Fothergill AW, McCarthy DI, Rinaldi MG, Graybill JR. In vitro activities of posaconazole, itraconazole, voriconazole, amphotericin B, and fluconazole against 37 clinical isolates of zygomycetes. Antimicrob Agents Chemother 2002; 46: 1581-1582.

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. Clin Infect Dis 2008; 46: 1813-1821.

Fridkin SK, Jarvis WR. Epidemiology of nosocomial fungal infections. Clin Microbiol Rev1996; 9: 499-511.

Scully C, de Almeida OP, Sposto MR. The deep mycoses in HIV infection. Oral Dis1997; 3: 200-207.

Tiraboschi I, Bravo M, Fernández N, Stecher D, Melero M, Lasala M. Mucormycosis. An emergent mycosis. Medicina (B Aires) 2012; 72: 23-27.

Beers MH, Robert BR(2004). Mucormycosis: In The Merck Manual of Diagnosis and Therapy. Whitehouse Station, NJ: Merck Research Laboratories.

Chayakulkeeree M, Ghannoum MA, Perfect JR. Zygomycosis: the re-emerging fungal infection. Eur J Clin Microbiol Infect Dis 2006; 25: 215-229.

Roden MM, Zaoutis TE, Buchanan WL, Knudsen TA, Sarkisova TA, Schaufele RL, et al. Epidemiology and outcome of zygomycosis: a review of 929 reported cases. Clin Infect Dis 2005; 41: 634-653.

Badiee P, Alborzi A, Karimi M, Pourabbas B, Haddadi P, Mardaneh J, et al. Diagnostic potential of nested PCR, galactomannan EIA, and beta-D-glucan for invasive aspergillosis in pediatric patients. J Infect Dev Ctries 2012; 13: 352-357.

Badiee P, Alborzi A. Detection of Aspergillus species in bone marrow transplant patients. J Infect Dev Ctries 2010; 4: 511-516.

Badiee P, Alborzi A, Vojdani R, Shakiba E, Rasouli M, Ravanfar P, et al. Early diagnosis of systemic candidiasis in bone marrow transplant recipients. Exp Clin Transplant 2010; 8: 98-103.

Lennette E, Balows A (1985). Manual of Clinical Microbiology: American Society for Microbiology. Washington DC.

Nosari A, Anghilieri M, Carrafiello G, Guffanti C, Marbello L, Montillo M, et al. Utility of percutaneous lung biopsy for diagnosing filamentous fungal infections in hematologic malignancies. Haematologica 2003; 88:1405-1409.

Badiee P, Alborzi A, Joukar M. Molecular assay to detect nosocomial fungal infections in intensive care units. Eur J Intern Med 2011; 22: 611-615.

Ribes JA, Vanover-Sams CL, Baker DJ. Zygomycetes in human disease. Clin Microbiol Rev 2000; 13: 236-301.

Roberts GD (2012). Clinical Mycology-Direct examination: zygomycetes. Mayo Foundation for Medical Education and Research.

Rickerts V, Just-Nübling G, Konrad F, Kern J, Lambrecht E, Böhme A, et al. Diagnosis of invasive aspergillosis and mucormycosis in immunocompromised patients by seminested PCR assay of tissue samples. Eur J Clin Microbiol Infect Dis 2006; 25: 8-13.

Gonzalez CE, Rinaldi MG, Sugar AM. Zygomycosis.Infect Dis Clin North Am 2002; 16: 895-914.

Frater JL, Hall GS, Procop GW. Histologic features of zygomycosis: emphasis on perineural invasion and fungal morphology. Arch Pathol Lab Med 2001; 125:375-378.

Kontoyiannis DP, Wessel VC, Bodey GP, Rolston KV.Zygomycosis in the 1990s in a tertiary-care cancer center. Clin Infect Dis 2000; 30: 851-856.

Spellberg B, Edwards J Jr, Ibrahim A. Novel perspectives on mucormycosis: pathophysiology, presentation, and management. Clin Microbiol Rev 2005; 18: 556-569.

Tarrand JJ, Han XY, Kontoyiannis DP, May GS.Aspergillus hyphae in infected tissue: evidence of physiologic adaptation and effect on culture recovery. J Clin Microbiol 2005; 43: 382-386.

Bialek R, Fischer J, Feucht A, Najvar LK, Dietz K, Knobloch J, et al. Diagnosis and monitoring of murine histoplasmosis by a nested PCR assay. J Clin Microbiol 2001; 39: 1506-1509.

Hata DJ, Buckwalter SP, Pritt BS, Roberts GD, Wengenack NL. Real-time PCR method for detection of zygomycetes. J Clin Microbiol 2008; 46: 2353-2358.

Bonin S, Petrera F, Niccolini B, Stanta G. PCR analysis in archival postmortem tissues. Mol Pathol 2003; 56:184-186.

Files
IssueVol 5 No 4 (2013) QRcode
SectionArticles
Keywords
18S rDNA Invasive fungal infection Mucoracea Semi-nested PCR

Rights and permissions
Creative Commons License This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
How to Cite
1.
Badiee P, Arastefar A, Jafarian H. Comparison of histopathological analysis, culture and polymerase chain reaction assays to detect mucormycosis in biopsy and blood specimens. Iran J Microbiol. 1;5(4):406-410.