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oai:repisalud.isciii.es:20.500.12105/128842024-09-27T20:30:01Zcom_20.500.12105_2052com_20.500.12105_2051col_20.500.12105_19609

00925njm 22002777a 4500 dc Bernal-Martinez, Leticia author Herrera-Leon, Laura author Valero, Clara author de la Cruz, Paula author Ghimpu, Larisa author Mesa-Arango, Ana Cecilia author Santoni, Gabriela author Goterris, Lidia author Millán, Rosario author Buitrago, Maria Jose author 2021-04-27 Opportunistic fungal pneumonias (OFP) are the main cause of death in AIDS patients worldwide. Diagnosis of these infections is often late as tuberculosis (TB) is frequently the first suspicion. In addition, diagnostic tools have limitations and are unavailable in disadvantaged regions. To perform the differential diagnosis of the main fungi causing OFP in AIDS patients (Histoplasma capsulatum, Cryptococcus neoformans/C. gattii and Pneumocystis jirovecii) vs. the Mycobacterium tuberculosis complex (MTBC), two new assays were developed: (i) a multiplex real-time PCR (MRT-PCR) and (ii) a simple and cost-effective method based on real-time PCR and the analysis of melting curves after amplification (MC-PCR). Both of the techniques were optimized and standardized "in vitro", showing a suitable reproducibility (CV ranged between 1.84 and 3.81% and 1.41 and 4.83%, respectively), a 100% specificity and detection limits between 20 and 2 fg of genomic DNA per 20 µL of reaction. A validation study was performed by retrospectively using 42 clinical samples from 37 patients with proven fungal infection or TB, and 33 controls. The overall sensitivity for the MRT-PCR assay and the MC-PCR assay was 88% and 90.4%, respectively. Both techniques were fast, sensitive and reproducible, allowing for the detection of these pathogens and the performance of a differential diagnosis. J Fungi (Basel) . 2021 Apr 27;7(5):336. http://hdl.handle.net/20.500.12105/12884 33925404 10.3390/jof7050336 2309-608X Journal of Fungi (Basel, Switzerland) Differential diagnosis Fungal pneumonia Tuberculosis Differential Diagnosis of Fungal Pneumonias vs. Tuberculosis in AIDS Patients by Using Two New Molecular Methods.