Rapid detection and identification of mucormycetes in bronchoalveolar lavage samples from immunocompromised patients with pulmonary infiltrates by use of high-resolution melt analysis

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Authors

LENGEROVÁ Martina RÁČIL Zdeněk HRNČÍŘOVÁ Kristýna KOCMANOVÁ Iva VOLFOVÁ Pavlína ŘÍČNÁ Dita BEJDÁK Petr MOULIS Mojmír PAVLOVSKÝ Zdeněk WEINBERGEROVÁ Barbora TOSKOVA Martina MAYER Jiří

Year of publication 2014
Type Article in Periodical
Magazine / Source Journal of Clinical Microbiology
MU Faculty or unit

Faculty of Medicine

Citation
Doi http://dx.doi.org/10.1128/JCM.00637-14
Field Oncology and hematology
Keywords Mucomycetes; PCR; fungal DNA; hematological malignancies
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Description Rapid differential diagnostics of pulmonary infiltrates suspected of invasive fungal disease in an immunocompromised host and early initiation of effective antifungal therapy are crucial for patient outcomes. There are no serological tests available to detect mucormycetes; therefore, PCR-based methods are highly suitable. We validated our previously published PCR followed by high-resolution melt analysis (PCR/HRMA) to detect Rhizopus spp., Rhizomucor pusillus, Lichtheimia corymbifera, and Mucor spp. in bronchoalveolar lavage (BAL) samples from immunocompromised patients who were at risk of invasive fungal disease. All PCR/HRMA-positive samples were retested using novel real-time quantitative PCR (RQ PCR) assays specific to the species identified. In total, between January 2009 and December 2012 we analyzed 99 BAL samples from 86 patients with pulmonary abnormalities using PCR/HRMA. Ninety (91%) BAL samples were negative, and 9 (9%) samples were positive. The sensitivity and specificity of PCR/HRMA were 100% and 93%, respectively. By combining the positive results of PCR/HRMA with positive RQ PCR results, the specificity was raised to 98%. PCR/HRMA, due to its high negative predictive value (99%), represents a fast and reliable tool for routine BAL sample screening for the differential diagnosis of pulmonary infiltrates in immunocompromised patients for the four most clinically important mucormycetes.
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