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A PCR-Based Approach for Early Diagnosis of Head and Neck Aspergillosis: A Pilot Study
Thaís Ellen Chaves Gomes1, Victor Coutinho Bastos2, Douglas Boniek3
1Department of Oral Surgery and Pathology, School of Dentistry, Universidade Federal de Minas Gerais, Belo Horizonte 31270-901, Brazil.
Background:
Aspergillosis is a fungal disease caused by the inhalation of fungal spores of the genus Aspergillus spp. This fungus mainly affects the lungs but can spread and infect the maxillofacial region through the bloodstream or inoculation of the fungus after extraction or endodontic treatment, especially in the upper posterior teeth. The disease has nonspecific clinical manifestations that hinder its early diagnosis. Although the Polymerase Chain Reaction (PCR) technique holds promise as a diagnostic tool for aspergillosis, anatomopathological analysis services do not routinely adopt this method.
Objectives:
Therefore, the present study aimed to evaluate the applicability of PCR and standardise the techniques of preparation of biological samples for the detection of the three species: Aspergillus niger, Aspergillus fumigatus and Aspergillus flavus. Methods: Six samples of formalin-fixed, paraffin-embedded tissue (FFPE) with a histopathological diagnosis suggestive of aspergillosis were investigated using PCR. As a positive control for the PCR reaction, morphologically and genetically characterized cultures were used, with their sequences deposited at NCBI under accession codes MW837777 (A. fumigatus) and MW837779 (A. niger). The A. flavus culture used is reference RC 2053.
Results:
Four of the six samples evaluated were positive for Aspergillus spp., of which one was co-infected with A. fumigatus and A. flavus species, while two others were positive only for A. flavus, and one sample was positive only for A. fumigatus.
Conclusions:
These findings suggest that PCR can be used as an auxiliary method for diagnosing aspergillosis. However, this was a pilot study, and expansion of the sample size and the evaluation of PCR in comparison with other diagnostic tests for aspergillosis are essential to determine the accuracy of the method.
Insights
Polymerase Chain Reaction (PCR) shows promise for diagnosing aspergillosis, a fungal infection. This pilot study demonstrated PCR
Area of Science:
- Medical Mycology
- Diagnostic Microbiology
- Infectious Diseases
Background:
- Aspergillosis is a fungal infection caused by Aspergillus spores, primarily affecting the lungs but potentially spreading to the maxillofacial region.
- Nonspecific clinical signs often delay diagnosis, despite the potential of Polymerase Chain Reaction (PCR) as a diagnostic tool.
- Current anatomopathological services do not routinely employ PCR for aspergillosis diagnosis.
Purpose of the Study:
- To assess the applicability of Polymerase Chain Reaction (PCR) for detecting Aspergillus species in clinical samples.
- To standardize sample preparation techniques for PCR-based aspergillosis diagnosis.
- To identify specific Aspergillus species, including Aspergillus niger, Aspergillus fumigatus, and Aspergillus flavus.
Main Methods:
- Investigated six formalin-fixed, paraffin-embedded (FFPE) tissue samples with suspected aspergillosis using PCR.
- Utilized characterized cultures of A. fumigatus and A. niger with NCBI accession numbers, and a reference culture for A. flavus as positive controls.
- Employed standardized sample preparation techniques for PCR analysis.
Main Results:
- PCR detected Aspergillus species in four out of six evaluated samples.
- One sample showed co-infection with Aspergillus fumigatus and Aspergillus flavus.
- Two samples tested positive for Aspergillus flavus only, and one sample tested positive for Aspergillus fumigatus only.
Conclusions:
- Polymerase Chain Reaction (PCR) can serve as an effective auxiliary method for diagnosing aspergillosis.
- This pilot study highlights the need for larger sample sizes and comparative evaluations with other diagnostic methods to confirm PCR's accuracy.
- Further research is essential to establish PCR as a routine diagnostic tool for aspergillosis.
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