Validation of a loop-mediated isothermal amplification assay for rapid diagnosis of invasive pneumococcal disease
Fecha de publicación
2020-03-24ISSN
2235-2988
Resumen
Current molecular PCR-based techniques used for detecting Streptococcus pneumoniae, the causative pathogen of invasive pneumococcal disease (IPD), are accurate but have a run time of several hours. We aimed to develop and validate a novel real-time loop mediated amplification (LAMP) assay for rapid detection of pneumococcus in normally sterile samples with accuracy comparable to a gold standard real-time PCR. Conserved regions of lytA were used for the design of the LAMP test. Analytical validation included assessment of linearity, limit of detection (LOD), intra-assay and inter-assay precision and analytical specificity, which was evaluated by using reference strain S. pneumoniae R6 and a quality control panel. Clinical performance was assessed on all samples collected from children with suspicion of IPD attended in Hospital Sant Joan de Deu (Barcelona, Spain) during the period April-September 2015. Fresh samples were analyzed after DNA extraction. The following values of analytical parameters were determined: linearity within the range 108-104 copies/mL; limit of detection, 5·103 copies/mL; intra- and inter-assay precision measured by mean coefficient of variance, 3.61 and 6.59%; analytical specificity, 9/9 pathogens similar to S. pneumoniae and 14/14 strains of different S. pneumoniae serotypes correctly identified as negative and positive results, respectively. Diagnostic sensitivity and specificity values were 100.0 and 99.3%. Median time of DNA amplification was 15 min. The new LAMP assay showed to have similar accuracy as PCR while being 5-fold faster and could become a useful diagnostic tool for early diagnosis of IPD.
Tipo de documento
Artículo
Versión del documento
Versión publicada
Lengua
English
Materias (CDU)
61 - Medicina
Palabras clave
Pneumococs
Microorganismes patògens
Antígens
Neumococos
Microorganismos
Antígenos
Pneumococcus
Pathogenic microorganisms
Antigens
Páginas
7
Publicado por
Frontiers Media
Colección
10; 115
Publicado en
Frontiers in Cellular and Infection Microbiology
Citación
de Paz, Héctor David Brotons, Pedro Esteva, Cristina [et al.]. Validation of a loop-mediated isothermal amplification assay for rapid diagnosis of invasive pneumococcal disease. Frontiers in Cellular and Infection Microbiology, 2020, 10(115), p. 1-7. Disponible en: <https://www.frontiersin.org/articles/10.3389/fcimb.2020.00115/full>. Fecha de acceso: 8 feb. 2020. DOI: 10.3389/fcimb.2020.00115
Número del acuerdo de la subvención
info:eu-repo/grantAgreement/EC/FP7/606488
Este ítem aparece en la(s) siguiente(s) colección(ones)
- Ciències de la Salut [740]
Derechos
© 2020 de Paz, Brotons, Esteva and Muñoz-Almagro. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
Excepto si se señala otra cosa, la licencia del ítem se describe como https://creativecommons.org/licenses/by/4.0/