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Assessment of a Loop-Mediated Isothermal Amplification (LAMP) Assay for the Rapid Detection of Pathogenic Bacteria from Respiratory Samples in Patients with Hospital-Acquired Pneumonia

Rapid identification of the causative agent of hospital-acquired pneumonia (HAP) will allow an earlier administration of a more appropriate antibiotic and could improve the outcome of these patients. The aim of this study was to develop a rapid protocol to identify the main microorganisms involved i...

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Autores principales: Vergara, Andrea, Boutal, Hervé, Ceccato, Adrián, López, Míriam, Cruells, Adrià, Bueno-Freire, Leticia, Moreno-Morales, Javier, Puig de la Bellacasa, Jorge, Castro, Pedro, Torres, Antoni, Marco, Francesc, Casals-Pascual, Climent, Vila, Jordi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7022425/
https://www.ncbi.nlm.nih.gov/pubmed/31940771
http://dx.doi.org/10.3390/microorganisms8010103
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author Vergara, Andrea
Boutal, Hervé
Ceccato, Adrián
López, Míriam
Cruells, Adrià
Bueno-Freire, Leticia
Moreno-Morales, Javier
Puig de la Bellacasa, Jorge
Castro, Pedro
Torres, Antoni
Marco, Francesc
Casals-Pascual, Climent
Vila, Jordi
author_facet Vergara, Andrea
Boutal, Hervé
Ceccato, Adrián
López, Míriam
Cruells, Adrià
Bueno-Freire, Leticia
Moreno-Morales, Javier
Puig de la Bellacasa, Jorge
Castro, Pedro
Torres, Antoni
Marco, Francesc
Casals-Pascual, Climent
Vila, Jordi
author_sort Vergara, Andrea
collection PubMed
description Rapid identification of the causative agent of hospital-acquired pneumonia (HAP) will allow an earlier administration of a more appropriate antibiotic and could improve the outcome of these patients. The aim of this study was to develop a rapid protocol to identify the main microorganisms involved in HAP by loop-mediated isothermal amplification (LAMP) directly from respiratory samples. First of all, a rapid procedure (<30 min) to extract the DNA from bronchoalveolar lavage (BAL), endotracheal aspirate (EA) or bronchoaspirate (BAS) was set up. A specific LAMP for Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Stenotrophomonas maltophilia and Acinetobacter baumannii was performed with the extracted solution at 65 °C for 30–40 min. Overall, 58 positive BAL and 83 EA/BAS samples were tested. The limits of detection varied according to the microorganism detected. Validation of the LAMP assay with BAL samples showed that the assay was 100% specific and 86.3% sensitive (positive predictive value of 100% and a negative predictive value of 50%) compared with culture. Meanwhile for BAS/EA samples, the assay rendered the following statistical parameters: 100% specificity, 94.6% sensitivity, 100% positive predictive value and 69.2% negative predictive value. The turnaround time including sample preparation and LAMP was circa 1 h. LAMP method may be used to detect the most frequent bacteria causing HAP. It is a simple, cheap, sensitive, specific and rapid assay.
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spelling pubmed-70224252020-03-09 Assessment of a Loop-Mediated Isothermal Amplification (LAMP) Assay for the Rapid Detection of Pathogenic Bacteria from Respiratory Samples in Patients with Hospital-Acquired Pneumonia Vergara, Andrea Boutal, Hervé Ceccato, Adrián López, Míriam Cruells, Adrià Bueno-Freire, Leticia Moreno-Morales, Javier Puig de la Bellacasa, Jorge Castro, Pedro Torres, Antoni Marco, Francesc Casals-Pascual, Climent Vila, Jordi Microorganisms Article Rapid identification of the causative agent of hospital-acquired pneumonia (HAP) will allow an earlier administration of a more appropriate antibiotic and could improve the outcome of these patients. The aim of this study was to develop a rapid protocol to identify the main microorganisms involved in HAP by loop-mediated isothermal amplification (LAMP) directly from respiratory samples. First of all, a rapid procedure (<30 min) to extract the DNA from bronchoalveolar lavage (BAL), endotracheal aspirate (EA) or bronchoaspirate (BAS) was set up. A specific LAMP for Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Stenotrophomonas maltophilia and Acinetobacter baumannii was performed with the extracted solution at 65 °C for 30–40 min. Overall, 58 positive BAL and 83 EA/BAS samples were tested. The limits of detection varied according to the microorganism detected. Validation of the LAMP assay with BAL samples showed that the assay was 100% specific and 86.3% sensitive (positive predictive value of 100% and a negative predictive value of 50%) compared with culture. Meanwhile for BAS/EA samples, the assay rendered the following statistical parameters: 100% specificity, 94.6% sensitivity, 100% positive predictive value and 69.2% negative predictive value. The turnaround time including sample preparation and LAMP was circa 1 h. LAMP method may be used to detect the most frequent bacteria causing HAP. It is a simple, cheap, sensitive, specific and rapid assay. MDPI 2020-01-11 /pmc/articles/PMC7022425/ /pubmed/31940771 http://dx.doi.org/10.3390/microorganisms8010103 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Vergara, Andrea
Boutal, Hervé
Ceccato, Adrián
López, Míriam
Cruells, Adrià
Bueno-Freire, Leticia
Moreno-Morales, Javier
Puig de la Bellacasa, Jorge
Castro, Pedro
Torres, Antoni
Marco, Francesc
Casals-Pascual, Climent
Vila, Jordi
Assessment of a Loop-Mediated Isothermal Amplification (LAMP) Assay for the Rapid Detection of Pathogenic Bacteria from Respiratory Samples in Patients with Hospital-Acquired Pneumonia
title Assessment of a Loop-Mediated Isothermal Amplification (LAMP) Assay for the Rapid Detection of Pathogenic Bacteria from Respiratory Samples in Patients with Hospital-Acquired Pneumonia
title_full Assessment of a Loop-Mediated Isothermal Amplification (LAMP) Assay for the Rapid Detection of Pathogenic Bacteria from Respiratory Samples in Patients with Hospital-Acquired Pneumonia
title_fullStr Assessment of a Loop-Mediated Isothermal Amplification (LAMP) Assay for the Rapid Detection of Pathogenic Bacteria from Respiratory Samples in Patients with Hospital-Acquired Pneumonia
title_full_unstemmed Assessment of a Loop-Mediated Isothermal Amplification (LAMP) Assay for the Rapid Detection of Pathogenic Bacteria from Respiratory Samples in Patients with Hospital-Acquired Pneumonia
title_short Assessment of a Loop-Mediated Isothermal Amplification (LAMP) Assay for the Rapid Detection of Pathogenic Bacteria from Respiratory Samples in Patients with Hospital-Acquired Pneumonia
title_sort assessment of a loop-mediated isothermal amplification (lamp) assay for the rapid detection of pathogenic bacteria from respiratory samples in patients with hospital-acquired pneumonia
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7022425/
https://www.ncbi.nlm.nih.gov/pubmed/31940771
http://dx.doi.org/10.3390/microorganisms8010103
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