Identification of pathogenic Aspergillus species by a PCR-restriction enzyme method

• Published in: Journal of medical microbiology Vol. 56; no. 11; pp. 1568 - 1570
• Main Authors: MIRHENDI, Hossein, DIBA, Kambiz, KORDBACHEH, Parivash, JALALIZAND, Nilufar, MAKIMURA, Kuichi
• Format: Journal Article
• Language: English
• Published: Reading Society for General Microbiology 01.11.2007
• Subjects: Aspergillosis - diagnosis; Aspergillosis - microbiology; Aspergillus - classification; Aspergillus - genetics; Aspergillus - isolation & purification; Aspergillus flavus; Aspergillus fumigatus; Aspergillus nidulans; Aspergillus niger; Aspergillus terreus; Aspergillus ustus; Biological and medical sciences; DNA Fingerprinting - methods; DNA, Fungal - genetics; DNA, Ribosomal Spacer - genetics; Fundamental and applied biological sciences. Psychology; Humans; Microbiology; Miscellaneous; Mycology; Polymerase Chain Reaction - methods; Polymorphism, Restriction Fragment Length; Fungi; Pathogenicity; Polymerase chain reaction; Aspergillus; Enzyme; Identification; Fungi Imperfecti; Method
• ISSN: 0022-2615; 1473-5644
• DOI: 10.1099/jmm.0.47319-0

Aspergillus fumigatus remains the most frequent cause of invasive aspergillosis; however, other species, including Aspergillus flavus, Aspergillus terreus, Aspergillus niger, Aspergillus nidulans and Aspergillus ustus have been reported to cause human infection (Henry et al., 2000). Rapid and accurate identification of Aspergillus species is necessary for successful clinical management of infection and for epidemiological purposes. Identification of Aspergillus species based on morphological methods requires adequate growth time for evaluation of colony characteristics and microscopic features. A culture time of 5 days or more is generally required for the development of anamorphic forms of Aspergillus. Failure to form conidia on ordinary culture media may require colonies to be further subcultured on specialized media to induce spore formation (Henry et al., 2000; Hinrikson et al., 2005). In addition, morphology tests are usually labour intensive and need expert mycology personnel. Due to these limitations, various molecular approaches have been used for the identification of Aspergillus species isolated from clinical samples, including PCR amplification of targets followed by either fragment length analysis or DNA probe hybridization or sequence analysis (Hinrikson et al., 2005). The aim of this study was to compare the internal transcribed spacer 1 (ITS1)-ITS2 nucleotide sequences of common Aspergillus species and design a PCR-RFLP profile for differentiation of the most medically important Aspergillus species.