Droplet Digital PCR for <i>Acinetobacter baumannii</i> Diagnosis in Bronchoalveolar Lavage Samples from Patients with Ventilator-Associated Pneumonia
Mirna Giselle Moreira,
Anna Gabriella Guimarães Oliveira,
Ihtisham Ul Haq,
Tatiana Flávia Pinheiro de Oliveira,
Wadi B. Alonazi,
Antônio Augusto Fonseca Júnior,
Vandack Alencar Nobre Junior,
Simone Gonçalves dos Santos
Affiliations
Mirna Giselle Moreira
Departament of Microbiology, Institute of Biological Sciences, Federal University of Minas Gerais, Av. Pres. Antônio Carlos 6627, Pampulha, Belo Horizonte 31270-901, Minas Gerais, Brazil
Anna Gabriella Guimarães Oliveira
Departament of Microbiology, Institute of Biological Sciences, Federal University of Minas Gerais, Av. Pres. Antônio Carlos 6627, Pampulha, Belo Horizonte 31270-901, Minas Gerais, Brazil
Ihtisham Ul Haq
Department of Physical Chemistry and Technology of Polymers, Silesian University of Technology, M. Strzody 9, 44-100 Gliwice, Poland
Tatiana Flávia Pinheiro de Oliveira
Federal Agricultural Defense Laboratory of Minas Gerais, Av. Rômulo Joviano s/n, Centro, Pedro Leopoldo 33600-000, Minas Gerais, Brazil
Wadi B. Alonazi
Health Administration Department, College of Business Administration, King Saud University, Riyadh 11421, Saudi Arabia
Antônio Augusto Fonseca Júnior
Federal Agricultural Defense Laboratory of Minas Gerais, Av. Rômulo Joviano s/n, Centro, Pedro Leopoldo 33600-000, Minas Gerais, Brazil
Vandack Alencar Nobre Junior
Interdisciplinary Center for Research in Intensive Care Medicine (NIIMI), Faculty of Medicine, Federal University of Minas Gerais, Av. Prof. Alfredo Balena 110, Santa Efigênia, Belo Horizonte 30130-100, Minas Gerais, Brazil
Simone Gonçalves dos Santos
Departament of Microbiology, Institute of Biological Sciences, Federal University of Minas Gerais, Av. Pres. Antônio Carlos 6627, Pampulha, Belo Horizonte 31270-901, Minas Gerais, Brazil
Advanced diagnostic technologies have made accurate and precise diagnosis of pathogens easy. Herein, we present a new diagnostic method, droplet digital PCR (ddPCR), to detect and quantify Acinetobacter baumannii in mini bronchoalveolar lavage (mini-BAL) samples. A. baumannii causes ventilator-associated pneumonia (VAP), a severe healthcare infection affecting patients’ lungs. VAP carries a high risk of morbidity and mortality, making its timely diagnosis crucial for prompt and effective management. Methodology. The assay performance was evaluated by comparing colonization data, quantitative culture results, and different generations of PCR (traditional PCR and Real-Time PCR—qPCR Taqman® and SYBR® Green). The ddPCR and qPCR Taqman® prove to be more sensitive than other molecular techniques. Reasonable analytical specificity was obtained with ddPCR, qPCR TaqMan®, and conventional PCR. However, qPCR SYBR® Green technology presented a low specificity, making the results questionable in clinical samples. DdPCR detected/quantified A. baumanni in more clinical samples than other methods (38.64% of the total samples). This emerging ddPCR technology offers promising advantages such as detection by more patients and direct quantification of pathogens without calibration curves.
