Rapid detection and identification of mucormycetes in bronchoalveolar lavage samples from immunocompromised patients with pulmonary infiltrates by use of high-resolution melt analysis.
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.
Department of Internal Medicine-Hematology and Oncology, University Hospital Brno, Brno, Czech Republic Department of Internal Medicine-Hematology and Oncology, Faculty of Medicine, Masaryk University, Brno, Czech Republic CEITEC-Central European Institute of Technology, Masaryk University, Brno, Czech Republic mlengerova@fnbrno.cz.
Martina
M
Lengerova
Department of Internal Medicine-Hematology and Oncology, University Hospital Brno, Brno, Czech Republic Department of Internal Medicine-Hematology and Oncology, Faculty of Medicine, Masaryk University, Brno, Czech Republic CEITEC-Central European Institute of Technology, Masaryk University, Brno, Czech Republic.
Zdenek
Z
Racil
Department of Internal Medicine-Hematology and Oncology, University Hospital Brno, Brno, Czech Republic.
Kristyna
K
Hrncirova
Department of Clinical Microbiology, University Hospital Brno, Brno, Czech Republic.
Iva
I
Kocmanova
Department of Internal Medicine-Hematology and Oncology, University Hospital Brno, Brno, Czech Republic.
Pavlina
P
Volfova
Department of Internal Medicine-Hematology and Oncology, University Hospital Brno, Brno, Czech Republic.
Dita
D
Ricna
Department of Internal Medicine-Hematology and Oncology, University Hospital Brno, Brno, Czech Republic.
Petr
P
Bejdak
Department of Pathology, University Hospital Brno, Brno, Czech Republic.
Mojmir
M
Moulis
Department of Pathology, University Hospital Brno, Brno, Czech Republic.
Zdenek
Z
Pavlovsky
Department of Internal Medicine-Hematology and Oncology, University Hospital Brno, Brno, Czech Republic.
Barbora
B
Weinbergerova
Department of Internal Medicine-Hematology and Oncology, University Hospital Brno, Brno, Czech Republic.
Martina
M
Toskova
Department of Internal Medicine-Hematology and Oncology, University Hospital Brno, Brno, Czech Republic Department of Internal Medicine-Hematology and Oncology, Faculty of Medicine, Masaryk University, Brno, Czech Republic CEITEC-Central European Institute of Technology, Masaryk University, Brno, Czech Republic.
Jiri
J
Mayer
D001992:Bronchoalveolar Lavage Fluid / Q000382:microbiology*; D005260:Female; D006801:Humans; D016867:Immunocompromised Host; D008172:Lung Diseases, Fungal / Q000175:diagnosis* / Q000382:microbiology; D008297:Male; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D009090:Mucorales / Q000145:classification* / Q000302:isolation & purification*; D009091:Mucormycosis / Q000175:diagnosis* / Q000382:microbiology; D016133:Polymerase Chain Reaction / Q000379:methods*; D011237:Predictive Value of Tests; D012680:Sensitivity and Specificity; D013997:Time Factors; D044366:Transition Temperature