Evaluation of nested PCR in diagnosis of fungal rhinosinusitis
Abstract
Background and Objective: Given the importance of rapid diagnosis for fungal rhinosinusitis, this study aimed to evaluate the use of nested PCR to identify Aspergillus and Mucor species in clinical samples from patients with suspected fungal rhinosinusitis.
Methods: Functional endoscopic sinus surgery specimens were collected from 98 patients with rhinosinusitis from 2012 to 2013. All samples were ground and cultured on sabouraud dextrose agar. The isolated fungi were identified based on their macroscopic and microscopic features. Fungal DNA was extracted from the tissue samples and nested PCR was performed with two sets of primers for Mucor and Aspergillus.
Results: Direct microscopic showed that 5.1% contained fungal components and 9.2% exhibited growth of fungi in culture. The most common agents isolated were Aspergillus fumigatus (n= 3 ), Aspergillus flavus (n=2), Penicillium sp (n=3) and Alternaria sp. (n=1). Mucor sp. was identified in the pathology smear from 1 patient. Positive results for fungal rhinosinusitis were obtained for a total of 10.2% by culture or pathology smear. Positive PCR results were obtained in 72 samples for Aspergillus and 31 samples for Mucor.
Conclusion: Our results suggest that endoscopic sinus surgery specimens are not suitable for nested PCR, probably because of the accumulation of fungi that contaminate the environmental air. This drawback is a limiting factor for diagnosis with nasal cavity specimens. Therefore, molecular methods and conventional culture techniques are helpful complementary diagnostic methods to detect fungal rhinosinusitis and determine appropriate management for these patients.
Benninger MS, Sedory Holzer SE, Lau J. Diagnosis and treatment of uncomplicated acute bacterial rhinosinusitis: Summary of the Agency for Health Care Policy and Research evidence-based report. Otolaryngol Head Neck Surg 2000; 122:1-7.
Lee LN, Bhattacharyya N. Regional and specialty variations in the treatment of chronic rhinosinusitis. Laryngoscope 2011; 121: 1092–7.
Pleis JR, Lucas JW, Ward BW. Summary health statistics for U.S. adults: National Health Interview Survey,2008. Vital Health Stat 2009; 10(242):1-157.
Badiee P, Jafarpour Z, Alborzi A, Haddadi P, Rasuli M, Kalani M. Orbital mucormycosis in an immunocompetent individual. Iran J Microbiol 2012;4:210-214.
Sohail MA, Al Khabori M, Hyder J, Verma A. Acute fulminant fungal sinusitis: Clinical presentation, radiological findings and treatment. Acta Trop 2001;80:177-85.
Das A, Bal A, Chakrabarti A, Panda N, Joshi K. Spectrum of fungal rhinosinusitis; histopathologist’s perspective. Histopathology 2009; 54: 854–859.
Catten MD, Murr AH, Goldstein JA, Mhatre AN, Lalwani AK. Detection of fungi in the nasal mucosa using polymerase chain reaction. Laryngoscope 2001;111: 399-403.
Badiee P, Nejabat M, Alborzi A, Keshavarz F, Shakiba E.Comparative study of Gram stain, potassium hydroxide smear, culture and nested PCR in the diagnosis of fungal keratitis. Ophthslmic Res 2010; 44:251-256.
Rickerts V, Just-Nübling G, Konrad F, Kern J, Lambrecht E, Böhme A, Jacobi V, 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.
Skladny H, Buchheidt D, Baust C, Krieg-Schneider F, Seifarth W, Leib-Mösch C, et al. Specific detection of Aspergillus species in blood and bronchoalveolar lavage samples of immunocompromised patients by two-step PCR. J Clin Microbiol 1999; 37:3865-3871.
Satish HS, Alokkan J. Clinical Study of Fungal Rhinosinusitis. IOSR Journal of Dental and Medical Sciences 2013; 5(4): 37-40.
Polzehl D, Weschta M, Podbielski A, Riechelmann H, Rimek D. Fungus culture and PCR in nasal lavage samples of patients with chronic rhinosinusitis. J Med Microbiol 2005; 54: 31–37.
Kordbacheh P, Zaini F, A Sabokbar, Borghei H, Safara M. Fungi as Causative Agent of Nasal Polyps in Tehran, Iran. Iran J Public Health 2006 35:53-57.
Granville L, Chirala M, Cernoch P, Ostrowski M, Truong LD. Fungal Sinusitis: Histologic Spectrum and Correlation with Culture. Hum Pathol 2004; 35:474-481.
Ferguson BJ. Fungus balls of the paranasal sinuses.Otolaryngol Clin North Am 2000; 33:389-398.
Menzes EA, Trindade ECP, Costa MM, Freire CCF, Márcio de Souza Cavalcante MS. Cunha FA. Airborne fungi isolated from Fortaleza City, State of Ceará, Brazil. Rev Inst Med Trop Sao Paulo 2004; 46:133-137.
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.
Rickerts V, Mousset S, Lambrecht E, Tintelnot K, Schwerdtfeger R, Presterl E, et al. Comparison of histopathological analysis, culture, and polymerase chain reaction assays to detect invasive mold infections from biopsy specimens. Clin Infect Dis 2007 15;44:1078-1083.
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 2013;5: 406-410.
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Issue | Vol 7 No 1 (2015) | |
Section | Original Article(s) | |
Keywords | ||
functional endoscopic sinus surgery rhinosinusitis nested PCR Aspergillus |
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