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...
| मुख्य लेखकों: | , , , , , , , , , , , , |
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| स्वरूप: | Journal article |
| भाषा: | English |
| प्रकाशित: |
MDPI
2020
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| _version_ | 1826306144267665408 |
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| author | 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 |
| author_facet | 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 |
| author_sort | Vergara, A |
| collection | OXFORD |
| 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. |
| first_indexed | 2024-03-07T06:43:28Z |
| format | Journal article |
| id | oxford-uuid:fa167cfb-c37b-46d6-a747-77feb5d9ce59 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-07T06:43:28Z |
| publishDate | 2020 |
| publisher | MDPI |
| record_format | dspace |
| spelling | oxford-uuid:fa167cfb-c37b-46d6-a747-77feb5d9ce592022-03-27T13:03:04ZAssessment of a Loop-Mediated Isothermal Amplification (LAMP) Assay for the Rapid Detection of Pathogenic Bacteria from Respiratory Samples in Patients with Hospital-Acquired PneumoniaJournal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:fa167cfb-c37b-46d6-a747-77feb5d9ce59EnglishSymplectic ElementsMDPI2020Vergara, ABoutal, HCeccato, ALópez, MCruells, ABueno-Freire, LMoreno-Morales, JPuig de la Bellacasa, JCastro, PTorres, AMarco, FCasals-Pascual, CVila, JRapid 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. |
| spellingShingle | 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 |
| 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 |
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