Paracoccidioidomycosis: Detection of Paracoccidioides brasiliensis´ genome in biological samples by quantitative chain reaction polymerase (qPCR)

• Published in: Microbial pathogenesis Vol. 121; pp. 359 - 362
• Main Authors: Rocha-Silva, Fabiana, Maria de Figueiredo, Sônia, Rutren La Santrer, Emanoelle Fernandes, Machado, Amanda Sanchez, Fernandes, Blenda, Assunção, Claudia Barbosa, Góes, Alfredo Miranda, Caligiorne, Rachel Basques
• Format: Journal Article
• Language: English
• Published: England Elsevier Ltd 01.08.2018
• Subjects: Brazil; Bronchoalveolar Lavage Fluid - microbiology; Diagnosis; DNA, Fungal - isolation & purification; Female; genes; Humans; Latin America; Latin America - epidemiology; Male; microbial detection; mycoses; P. lutzii; Paracoccidioides - genetics; Paracoccidioides - isolation & purification; Paracoccidioides brasiliensis; Paracoccidioidomycosis; Paracoccidioidomycosis - diagnosis; Paracoccidioidomycosis - epidemiology; Paracocciodioides brasiliensis; patients; Prevalence; Protein Pb27; public health; qPCR; Quantitative PCR; quantitative polymerase chain reaction; Real-Time Polymerase Chain Reaction; recombinant proteins; Reproducibility of Results; saprophytes; Sensitivity and Specificity; species identification; Brazil; Latin America; P. lutzii; Paracoccidioidomycosis; qPCR; Paracocciodioides brasiliensis; Diagnosis; Protein Pb27; Quantitative PCR
• ISSN: 0882-4010; 1096-1208; 1096-1208
• DOI: 10.1016/j.micpath.2018.05.039

Paracoccidioidomycosis (PCM) is a chronic mycosis caused by the saprobic and dimorphic species Paracoccidioides brasiliensis and P. lutzii. This disease is prevalent in Latin American countries. PCM appears as a relevant concern and challenge for the mycologists, since until now there is no a methodology suitable for an efficient and safe diagnosis and species identification. Thus, the present study aimed to validate a methodology for PCM´s diagnosis, using quantitative Polymerase Chain Reaction (qPCR) through target amplification of the gene encoding the recombinant protein Pb27, a common protein to the both species Paracoccidioides brasiliensis and P. lutzii. The experiments were performed in vitro to determine the specificity, efficiency and detection limit of qPCR assay, using specific primers and probe, which sequences were subject to a patent deposited in Brazilian CTIT, under the registration number: BR1020160078830. According to the results the technique showed sensitivity of 94% and specificity of 100%, demonstrating that it will be possible to develop a new fast and safe diagnostic PCM and can be standardized in order to present a low cost, accessible to the patient served by the public health system in Brazil and Latin America.