Differential Diagnosis of Fungal Pneumonias vs. Tuberculosis in AIDS Patients by Using Two New Molecular Methods

Journal of Fungi

Laura Herrera¹  Leticia Bernal-Martínez²  MaríaJosé Buitrago²  Clara Valero²  Paula de la Cruz²  Lidia Goterris²  AnaC. Mesa-Arango²  Larisa Ghimpu²  Gabriela Santoni²  Rosario Millán³ 

[1] Centro Nacional de Microbiología, Mycobacteria Unit, Instituto de Salud Carlos III, 28220 Madrid, Spain;Centro Nacional de Microbiología, Mycology Reference Laboratory, Instituto de Salud Carlos III, Majadahonda, 28220 Madrid, Spain;Department of Microbiology, Hospital Universitario Puerta de Hierro, 28220 Madrid, Spain;

关键词: differential diagnosis;    tuberculosis;    fungal pneumonia;   

DOI  :  10.3390/jof7050336

来源: DOAJ

【 摘 要 】

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.

【 授权许可】

Unknown