{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,19]],"date-time":"2026-10-19T20:51:12Z","timestamp":1792443072821,"version":"4.5.0"},"reference-count":43,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2018,2,9]],"date-time":"2018-02-09T00:00:00Z","timestamp":1518134400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Bacterial meningitis infection is a leading global health concern for which rapid and accurate diagnosis is essential to reduce associated morbidity and mortality. Loop-mediated isothermal amplification (LAMP) offers an effective low-cost diagnostic approach; however, multiplex LAMP is difficult to achieve, limiting its application. We have developed novel real-time multiplex LAMP technology, TEC-LAMP, using Tth endonuclease IV and a unique LAMP primer\/probe. This study evaluates the analytical specificity, limit of detection (LOD) and clinical application of an internally controlled multiplex TEC-LAMP assay for detection of leading bacterial meningitis pathogens: Streptococcus pneumoniae, Neisseria meningitidis and Haemophilus influenzae. Analytical specificities were established by testing 168 bacterial strains, and LODs were determined using Probit analysis. The TEC-LAMP assay was 100% specific, with LODs for S. pneumoniae, N. meningitidis and H. influenzae of 39.5, 17.3 and 25.9 genome copies per reaction, respectively. Clinical performance was evaluated by testing 65 archived PCR-positive samples. Compared to singleplex real-time PCR, the multiplex TEC-LAMP assay demonstrated diagnostic sensitivity and specificity of 92.3% and 100%, respectively. This is the first report of a single-tube internally controlled multiplex LAMP assay for bacterial meningitis pathogen detection, and the first report of Tth endonuclease IV incorporation into nucleic acid amplification diagnostic technology.<\/jats:p>","DOI":"10.3390\/ijms19020524","type":"journal-article","created":{"date-parts":[[2018,2,9]],"date-time":"2018-02-09T12:46:27Z","timestamp":1518180387000},"page":"524","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":39,"title":["Evaluation of an Internally Controlled Multiplex Tth Endonuclease Cleavage Loop-Mediated Isothermal Amplification (TEC-LAMP) Assay for the Detection of Bacterial Meningitis Pathogens"],"prefix":"10.3390","volume":"19","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-9420-8368","authenticated-orcid":false,"given":"Owen","family":"Higgins","sequence":"first","affiliation":[{"name":"Molecular Diagnostics Research Group, School of Natural Sciences and National Centre for Biomedical Engineering Science, National University of Ireland, Galway, Ireland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eoin","family":"Clancy","sequence":"additional","affiliation":[{"name":"Molecular Diagnostics Research Group, School of Natural Sciences and National Centre for Biomedical Engineering Science, National University of Ireland, Galway, Ireland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Martin","family":"Cormican","sequence":"additional","affiliation":[{"name":"School of Medicine, National University of Ireland, Galway and Galway University Hospital, Galway, Ireland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Teck","family":"Boo","sequence":"additional","affiliation":[{"name":"School of Medicine, National University of Ireland, Galway and Galway University Hospital, Galway, Ireland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Robert","family":"Cunney","sequence":"additional","affiliation":[{"name":"Irish Meningitis and Sepsis Reference Laboratory, Temple Street Children\u2019s University Hospital, Temple Street, Dublin, Ireland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4956-3299","authenticated-orcid":false,"given":"Terry","family":"Smith","sequence":"additional","affiliation":[{"name":"Molecular Diagnostics Research Group, School of Natural Sciences and National Centre for Biomedical Engineering Science, National University of Ireland, Galway, Ireland"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2018,2,9]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"467","DOI":"10.1128\/CMR.00070-09","article-title":"Epidemiology, diagnosis, and antimicrobial treatment of acute bacterial meningitis","volume":"23","author":"Brouwer","year":"2010","journal-title":"Clin. 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