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Diagnosis of mucormycosis using an intercalating dye-based quantitative PCR
Jeanne Bigot1, Alexandre Godmer2,3, Lysa Prudenté4
1Sorbonne Université, INSERM, Centre de Recherche Saint-Antoine, CRSA, APHP, Hôpital Saint Antoine, Laboratoire de Parasitologie Mycologie, 75012 Paris, France.
Medical Mycology
|February 21, 2022
Summary
A new quantitative PCR (qPCR) assay effectively detects a broad range of Mucorales species, aiding in the diagnosis of mucormycosis. This sensitive and specific molecular tool shows promise for clinical application in identifying this serious fungal infection.
Area of Science:
- Medical Mycology
- Molecular Diagnostics
- Infectious Diseases
Background:
- Mucormycosis is a serious fungal infection with high mortality.
- Accurate and timely diagnosis remains challenging.
- PCR-based methods offer a promising avenue for early detection.
Purpose of the Study:
- To develop and evaluate a single-tube, multiplex quantitative PCR (qPCR) assay for detecting diverse Mucorales species.
- To assess the analytical and clinical performance of the developed qPCR assay for mucormycosis diagnosis.
Main Methods:
- Design of degenerate primers targeting the rDNA operon for broad Mucorales detection.
- Utilized qPCR with an intercalating fluorescent dye.
- Analytical validation with 8 genera/11 species of Mucorales and 9 genera/14 species of non-Mucorales.
- Prospective testing on 364 clinical specimens from 166 at-risk patients.
Main Results:
- 100% sensitivity and specificity in analytical testing with a limit of detection of 3 rDNA copies/reaction.
- Positive qPCR results in all 7 proven/probable mucormycosis cases and 1 proven uncharacterized invasive mold infection.
- Sensitivity ranged from 73.33% to 100% and specificity from 98.10% to 100% in clinical specimens.
- 3 out of 7 possible mold invasive infections showed at least one positive qPCR test.
Conclusions:
- The developed pan-Mucorales qPCR assay demonstrates excellent analytical and clinical performance.
- This qPCR method is a valuable adjunctive tool for the challenging diagnosis of mucormycosis.
- The assay's ability to detect multiple species in a single reaction enhances diagnostic efficiency.
