Droplet Digital PCR for Acinetobacter baumannii 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 i...

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Published in:Antibiotics (Basel) 2024-09, Vol.13 (9), p.878
Main Authors: Giselle Moreira, Mirna, Guimarães Oliveira, Anna Gabriella, Ul Haq, Ihtisham, Pinheiro de Oliveira, Tatiana Flávia, Alonazi, Wadi B, Fonseca Júnior, Antônio Augusto, Nobre Junior, Vandack Alencar, Santos, Simone Gonçalves Dos
Format: Article
Language:English
Subjects:
Acinetobacter baumannii
Acinetobacter infections
Alveoli
Antimicrobial agents
Bacteria
Bacterial pneumonia
Bronchoalveolar lavage
Bronchus
Clean technology
Confidence intervals
Diagnosis
Diagnostic systems
droplet digital PCR
Droplets
Drug resistance
Drug resistance in microorganisms
Health aspects
Imipenem
Lavage
Microorganisms
Morbidity
Mortality
Nosocomial infections
Ostomy
Pathogens
Patients
Pneumonia
Polymerase chain reaction
real-time PCR
Staphylococcus infections
Ventilator-associated pneumonia
Ventilators
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Summary: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.
ISSN:2079-6382
2079-6382
DOI:10.3390/antibiotics13090878