RESEARCH PAPER
Genotypic discrimination of Aspergillus fumigatus strain from related species within section fumigati
 
More details
Hide details
1
Faculty of Science, Department of Biology, Basic and Industrial Microbiology Section, Ege University, Turkey
 
 
Corresponding author
Füsun B. Uçar   

Faculty of Science, Department of Biology, Basic and Industrial Microbiology Section, Ege University, Turkey
 
 
Ann Agric Environ Med. 2016;23(3):448-451
 
KEYWORDS
ABSTRACT
Introduction and objective:
The aim was to make an exact diagnosis of 20 strains using molecular biological methods which were isolated from the atmosphere of the inpatient rooms in the Oncology and other departments of the Ege University Medical Faculty Hospital, and identified as Aspergillus fumigatus through phenotypic tests, and to determine their antibiotic susceptibility patterns.

Material and Methods:
It was confirmed that the 20 phenotypically-identified A. fumigatus strains belonged to the section Fumigati after they were tested by the ITS-PCR method. Their sequence analysis was performed and the results sent to the NCBI GenBank, and their accession numbers were obtained. For their exact diagnosis at the species level, the β-tub (β-tubulin) and rodA (RodletA) genes were examined with the multiplex PCR. Anti-fungal susceptibility of the 20 strains was determined according to the M38-A2 standards.

Results:
As a result of ITS-PCR, it was confirmed that 19 of the 20 strains identified as A. fumigatus through the phenotypic methods belonged to the section Fumigati. However, after the detection of β-tub and rodA genes, all 20 strains were identified as A. fumigatus.

Conclusions:
Although the results of the phenotypic and molecular biological tests applied to filamentous fungi do not often overlap, in this study, the results obtained from the molecular analysis confirmed the results of the phenotypic tests. However, 1 of the 20 strains phenotypically-identified as A. fumigatus was identified as Penicillium spp. as a result of ITS-PCR and sequence analysis. On the other hand, the profile obtained from β-tub and rodA tests indicated that the strain was A. fumigatus. Based on these results, this strain is thought to belong to the Aspergilloides genus which has the features of both genera.

REFERENCES (21)
1.
Lasker BA. Evaluation of performance four genotypic methods for studying the genetic epidemiology of Aspergillus fumigatus isolates. J Clin Microbiol. 2002; 40(8): 2886–2892.
 
2.
Rakeman JL, Bui U, LaFe K, Chen Y, Honeycutt RJ, Cookson BT. Multilocus DNA sequence comparisons rapidly identify pathogenic molds. J Clin Microbiol. 2005; 43(7): 3324–3333.
 
3.
Henry T, Iwen PC, Hınrıcs SH. Identification of Aspergillus species using internal transcribed spacer region 1 and 2. J Clin Microbiol. 2000; 38(4): 1510–1515.
 
4.
Abaci O, Haliki-Uztan A, Ates M, Bacakoglu F. Detectıon of Aerıal Mıcrofungus Flora, Potentıal Opportunıst Infectıon Agent, ın the Intensıve Care Unıt of The Chest Dısease Department of a Hospıtal ın Izmır. 2012; 21 (2): 351–356..
 
5.
Montenegro G, Puch SS, Jewtuchowicz VM, Pinoni MV, Relloso S, Temporitti E, et al. Phenotypic and genotypic characterization of Aspergillus lentulus and Aspergillus fumigatus isolates in a patient with probable invasive aspergillosis. J Med Microbiol. 2009; 58(3): 391–395.
 
6.
Verweij PE, Meis JFGM, Sarfati J, Hoogkamp-Korstange JAA, Latge JP, – Melchers WJG. Genotypic characterization of sequential Aspergillus fumigatus isolates from patients with cystic fibrosis. J Clin Microbiol. 1996; 34(10): 2595–2597.
 
7.
Balajee SA, Gribskov JL, Hanley E, Nickle D, Marr KA. Aspergillus lentulus sp. nov., a new sibling species of A. fumigatus. Eukaryot Cell. 2005; 4(3): 625–632.
 
8.
Balajee SA, Nickle D, Varga J, Marr KA. Molecular Studies Reveal Frequent Misidentification of Aspergillus fumigatus by Morphotyping. Eukaryot Cell. 2006; 5(10): 1705–1712.
 
9.
Walsh TJ, Petraitis V, Petraitiene R, Field-Ridley A, Sutton D, Ghannoum M, et al. Experimental pulmonary aspergillosis due to Aspergillus terreus: pathogenesis and treatment of an emerging pathogen resistant to amphotericin B. J Infect Dis. 2003; 188(2): 305–309.
 
10.
Balajee SA, Houbraken J, Verweji PE, Hong SB, Yaghuchi T, Varga J, et al. Aspergillus species identification in the clinical setting. Stud Mycol. 2007; 59: 39–46.
 
11.
Glass NL, Donaldson GC. Development of primer sets designed for use with the PCR to amplify conserved genes from filamentous Ascomycetes. Appl Environ Microbiol. 1995; 61(4): 1323–1330.
 
12.
Hay RJ, Jones RM. New molecular tools in the diagnosis of superficial fungal infections. Clin Dermatol. 2010; 28(2): 190–196.
 
13.
Serrano R, Gusmão1 L, Amorim A, Araujo R. Rapid identification of Aspergillus fumigatus within the section Fumigati. BMC Microbiol. 2011; 11: 82.
 
14.
Klich MA. Identification of common Aspergillus species. AD Utrecht, The Netherlands, 2002.
 
15.
National Committee for Clinical Laboratory Standards. Reference method for broth dilution antifungal susceptibility testing of filamentous fungi. Approved standard M38-A. 2002, National Committee for Clinical Labora- tory Standards, Wayne, Pa.
 
16.
Liu D, Coloe S, Baird R, Pedersen J. Rapid mini-preparation of fungal DNA for PCR. J Clin Microbiol. 2000; 38(1): 471.
 
17.
Yalçın HT, Uçar FB. Isolation and characterization of cheese spoiler yeast isolated from Turkish White cheeses. Ann Microbiol. 2009; 59(3): 477–483.
 
18.
Balajee SA, Kano R, Baddley JW, Moser SA, Marr KA, Alexander BA, et al. Molecular identification of Aspergillus species collected for the transplant-associated infection surveillance network. J Clin Microbiol. 2009; 47(10): 3138–3141.
 
19.
Balajee SA, Gribskov J, Brandt M, Ito J, Fothergill A, Marr KA. Mistaken identity: Neosartorya pseudofischeri and its anamorph masquerading as Aspergillus fumigatus. J Clin Microbiol. 2005; 43(12): 5996–5999.
 
20.
Hong SB, Shin HD, Hong J, Frisvad JC, Nielsen PV, Varga J, et al. New taxa of Neosartorya and Aspergillus in Aspergillus section Fumigati. Antonie van Leeuwenhoek. 2008; 93(1–2): 87–98.
 
21.
Pıtt JI. A laboratory guide to common Penicillium species. 3nd ed. Food Science Australia, 2000.
 
eISSN:1898-2263
ISSN:1232-1966
Journals System - logo
Scroll to top