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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

1
Department of Clinical Microbiology, CDB, Hospital Clínic of Barcelona, University of Barcelona, 08036 Barcelona, Spain
2
Service de Pharmacologie et Immunoanalyse (SPI), CEA, INRA, Laboratoire d’Etudes et de Recherches en Immunonalyse, Université Paris-Saclay, F-91191 Gif-sur-Yvette, France
3
Pulmonary and Critical Care Unit, Respiratory Institute, Hospital Clinic, 08036 Barcelona, Spain
4
Biomedical Research Centre in Respiratory Diseases (CibeRes, CB06/06/0028), Institut d’Investigacions Biomèdiques Agust Pi i Sunyer (IDIBAPS), 08036 Barcelona, Spain
5
Department of Medicine, School of Medicine, University of Barcelona, 08036 Barcelona, Spain
6
ISGlobal, Hospital Clínic–Universitat de Barcelona, 08036 Barcelona, Spain
7
Medical Intensive Care Unit, Hospital Clínic, 08036 Barcelona, Spain
*
Author to whom correspondence should be addressed.
These authors contributed equally to this work.
Microorganisms 2020, 8(1), 103; https://doi.org/10.3390/microorganisms8010103
Received: 5 December 2019 / Revised: 24 December 2019 / Accepted: 8 January 2020 / Published: 11 January 2020
(This article belongs to the Section Medical Microbiology)
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. View Full-Text
Keywords: loop-mediated isothermal amplification (LAMP); hospital-acquired pneumonia (HAP); diagnostic techniques; respiratory system; critical care loop-mediated isothermal amplification (LAMP); hospital-acquired pneumonia (HAP); diagnostic techniques; respiratory system; critical care
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MDPI and ACS Style

Vergara, A.; Boutal, H.; Ceccato, A.; López, M.; Cruells, A.; Bueno-Freire, L.; Moreno-Morales, J.; Puig de la Bellacasa, J.; Castro, P.; Torres, A.; Marco, F.; Casals-Pascual, C.; Vila, J. 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. Microorganisms 2020, 8, 103. https://doi.org/10.3390/microorganisms8010103

AMA Style

Vergara A, Boutal H, Ceccato A, López M, Cruells A, Bueno-Freire L, Moreno-Morales J, Puig de la Bellacasa J, Castro P, Torres A, Marco F, Casals-Pascual C, Vila J. 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. Microorganisms. 2020; 8(1):103. https://doi.org/10.3390/microorganisms8010103

Chicago/Turabian Style

Vergara, Andrea, Hervé Boutal, Adrián Ceccato, Míriam López, Adrià Cruells, Leticia Bueno-Freire, Javier Moreno-Morales, Jorge Puig de la Bellacasa, Pedro Castro, Antoni Torres, Francesc Marco, Climent Casals-Pascual, and Jordi Vila. 2020. "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" Microorganisms 8, no. 1: 103. https://doi.org/10.3390/microorganisms8010103

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