Droplet Digital PCR for Diagnosis in Bronchoalveolar Lavage Samples from Patients with Ventilator-Associated Pneumonia.

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 in mini bronchoalveolar lavage (mini-BAL) samples. 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 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.