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Molecular identification of Aspergillus fumigatus complex from lung transplant recipients using multilocus sequencing
Ghulam Dhabaan1,2,3, Julianne Kus3,4, Deepali Kumar2
1Department of Microbiology, Sinai Health System/University Health Network, Toronto, Ontario, Canada.
Background:
Aspergillus infection causes significant morbidity and mortality among lung transplant recipients (LTRs). It is primarily caused by Aspergillus fumigatus. Other closely related species belonging to the section Fumigati have also been found. These cryptic species are often misidentified as A. fumigatus. Thus, we used multilocus sequencing analysis (MLSA) of the calmodulin, β-tubulin, and hydrophobin gene sequences to identify these species and to determine the frequency with which they occur among LTRs.
Methods:
A total of 81 A. fumigatus isolates were initially isolated from bronchoalveolar lavage fluid or sputum specimens collected from lung transplant patients. These isolates were then sub-cultured and genotyped using MLSA. Of these isolates, 53, 17, and 11 were isolated from double LTRs, single LTRs, and pre-LTRs, respectively.
Results:
All isolates (100%) carried DNA sequences identical to those of A. fumigatus reference strains and thus clustered in the same clade with A. fumigatus. Analysis of the MLSA data revealed that A. fumigatus species were the only species recovered in this population of LTRs. The MLSA results were consistent with those routinely obtained by conventional mycological procedures in the microbiology laboratory.
Conclusions:
A. fumigatus appears to be the primary causative agent of colonization or invasive aspergillosis among LTRs. No cryptic species were identified.
Insights
Aspergillus fumigatus is the main cause of aspergillosis in lung transplant recipients. Multilocus sequencing analysis confirmed no cryptic species were present in the studied patient population.
Area of Science:
- Medical Mycology
- Transplant Infectious Diseases
Background:
- Aspergillus infection poses a significant threat to lung transplant recipients (LTRs), often caused by Aspergillus fumigatus.
- Cryptic species closely related to A. fumigatus can be misidentified, complicating diagnosis and treatment.
Purpose of the Study:
- To accurately identify Aspergillus species causing infections in LTRs.
- To determine the prevalence of cryptic Aspergillus species in this patient group using multilocus sequencing analysis (MLSA).
Main Methods:
- Collected 81 Aspergillus isolates from bronchoalveolar lavage or sputum of LTRs.
- Genotyped isolates using MLSA with calmodulin, β-tubulin, and hydrophobin gene sequences.
Main Results:
- All 81 isolates showed DNA sequences identical to A. fumigatus reference strains.
- MLSA confirmed that only A. fumigatus species were present in the LTRs studied.
- Results aligned with conventional mycological identification methods.
Conclusions:
- A. fumigatus is the predominant pathogen responsible for aspergillosis in LTRs.
- No evidence of cryptic Aspergillus species was found in this cohort of lung transplant recipients.
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