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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
Author: Vergara, Andrea
Boutal, Hervé
Ceccato, Adrian
López, Miriam
Cruells, Adrián
Bueno, Leticia
Moreno-Morales, Javier
Puig de la Bellacasa, Jordi
Castro, Pedro
Torres Martí, Antoni
Marco Reverté, Francesc
Casals Pascual, Climent
Vila Estapé, Jordi
Keywords: Diagnòstic
Medicina intensiva
Aparell respiratori
Pneumònia adquirida a la comunitat
Critical care medicine
Respiratory organs
Community-acquired pneumonia
Issue Date: 11-Jan-2020
Publisher: MDPI
Abstract: Rapid identification of the causative agent of hospital-acquired pneumonia (HAP) will allowan earlier administration of a more appropriate antibiotic and could improve the outcome of thesepatients. The aim of this study was to develop a rapid protocol to identify the main microorganismsinvolved 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 forStaphylococcusaureus,Escherichia coli,Klebsiella pneumoniae, Pseudomonas aeruginosa,Stenotrophomonas maltophiliaandAcinetobacter baumanniiwas 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 themicroorganism 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 valueof 50%) compared with culture. Meanwhile for BAS/EA samples, the assay rendered the followingstatistical 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 circa1 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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It is part of: Microorganisms, 2020, vol. 8, num. 1, p. 103
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ISSN: 2076-2607
Appears in Collections:Articles publicats en revistes (Medicina)
Articles publicats en revistes (IDIBAPS: Institut d'investigacions Biomèdiques August Pi i Sunyer)
Articles publicats en revistes (ISGlobal)
Articles publicats en revistes (Fonaments Clínics)

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