{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,2]],"date-time":"2026-04-02T13:22:55Z","timestamp":1775136175559,"version":"3.50.1"},"reference-count":21,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2024,3,30]],"date-time":"2024-03-30T00:00:00Z","timestamp":1711756800000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2024,3,30]],"date-time":"2024-03-30T00:00:00Z","timestamp":1711756800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"name":"Vietnamese Ministry of National Defence","award":["364\/2020\/HD-NCKHCN"],"award-info":[{"award-number":["364\/2020\/HD-NCKHCN"]}]},{"name":"Vietnamese Ministry of National Defence","award":["364\/2020\/HD-NCKHCN"],"award-info":[{"award-number":["364\/2020\/HD-NCKHCN"]}]},{"name":"Vietnamese Ministry of National Defence","award":["364\/2020\/HD-NCKHCN"],"award-info":[{"award-number":["364\/2020\/HD-NCKHCN"]}]},{"name":"Vietnamese Ministry of National Defence","award":["364\/2020\/HD-NCKHCN"],"award-info":[{"award-number":["364\/2020\/HD-NCKHCN"]}]},{"name":"PAN-ASEAN Coalition for Epidemic and Outbreak Preparedness","award":["DAAD-57592343"],"award-info":[{"award-number":["DAAD-57592343"]}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Ann Clin Microbiol Antimicrob"],"abstract":"<jats:title>Abstract<\/jats:title><jats:sec>\n                <jats:title>Background<\/jats:title>\n                <jats:p><jats:italic>Neisseria meningitidis<\/jats:italic> can cause life-threatening meningococcal meningitis and meningococcemia. Old standard microbiological results from CSF\/blood cultures are time consuming. This study aimed to combine the sensitivity of loop-mediated isothermal nucleic acid amplification (LAMP) with the specificity of CRISPR\/Cas12a cleavage to demonstrate a reliable diagnostic assay for rapid detection of <jats:italic>N. meningitidis<\/jats:italic>.<\/jats:p>\n              <\/jats:sec><jats:sec>\n                <jats:title>Methods<\/jats:title>\n                <jats:p>A total of <jats:italic>n<\/jats:italic>\u2009=\u2009139 samples were collected from patients with suspected meningococcal disease and were used for evaluation. The extracted DNA was subjected to qualitative real-time PCR, targeting capsular transporter gene (<jats:italic>ctrA)<\/jats:italic> of <jats:italic>N. meningitidis<\/jats:italic>. LAMP-specific primer pairs, also targeting the <jats:italic>ctrA,<\/jats:italic> were designed and the LAMP products were subjected to CRISPR\/Cas12 cleavage reaction. the readout was on a lateral flow strip. Sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of LAMP-CRISPR\/Cas was compared with real-time PCR assays. The limit of detection (LOD) was established with serial dilutions of the target <jats:italic>N. meningitidis<\/jats:italic> DNA and calculated by Probit regression analysis.<\/jats:p>\n              <\/jats:sec><jats:sec>\n                <jats:title>Results<\/jats:title>\n                <jats:p>Six LAMP assay-specific primers were developed targeting the <jats:italic>ctrA<\/jats:italic> gene of <jats:italic>N. meningitidis<\/jats:italic>, which is conserved in all meningococcal serogroups. The LAMP primers did not amplify DNA from other bacterial DNA tested, showing 100% specificity. The use of 0.4\u00a0M betaine increased the sensitivity and stability of the reaction. LAMP-CRISPR\/Cas detected meningococcal serogroups (B, C, W). The assay showed no cross-reactivity and was specific for <jats:italic>N. meningitidis<\/jats:italic>. The LOD was 74 (95% CI: 47\u2013311) <jats:italic>N. meningitidis<\/jats:italic> copies. The LAMP-CRISPR\/Cas performed well compared to the gold standard. In the 139 samples from suspected patients, the sensitivity and specificity of the test were 91% and 99% respectively.<\/jats:p>\n              <\/jats:sec><jats:sec>\n                <jats:title>Conclusion<\/jats:title>\n                <jats:p>This developed and optimized method can complement for the available gold standard for the timely diagnosis of meningococcal meningitis and meningococcemia.<\/jats:p>\n              <\/jats:sec>","DOI":"10.1186\/s12941-024-00688-1","type":"journal-article","created":{"date-parts":[[2024,3,30]],"date-time":"2024-03-30T18:01:46Z","timestamp":1711821706000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":15,"title":["An isothermal CRISPR- based lateral flow assay for detection of Neisseria meningitidis"],"prefix":"10.1186","volume":"23","author":[{"given":"Dao Thi","family":"Huyen","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Julien","family":"Reboud","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Dao Thanh","family":"Quyen","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jonathan M.","family":"Cooper","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Thirumalaisamy P.","family":"Velavan","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ngo Tat","family":"Trung","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Le Huu","family":"Song","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2024,3,30]]},"reference":[{"key":"688_CR1","doi-asserted-by":"crossref","unstructured":"Chandran A et al. Long-term sequelae of childhood bacterial meningitis: an underappreciated problem. 2011;30(1):3\u20136.","DOI":"10.1097\/INF.0b013e3181ef25f7"},{"key":"688_CR2","doi-asserted-by":"crossref","unstructured":"Mazamay S et al. An overview of bacterial meningitis epidemics in Africa from 1928 to 2018 with a focus on epidemics outside-the-belt. 2021;21(1):1\u201313.","DOI":"10.1186\/s12879-021-06724-1"},{"key":"688_CR3","doi-asserted-by":"crossref","unstructured":"Aye AMM et al. Meningococcal disease surveillance in the Asia\u2013Pacific region (2020): the global meningococcal initiative. 2020;81(5):698\u2013711.","DOI":"10.1016\/j.jinf.2020.07.025"},{"key":"688_CR4","doi-asserted-by":"crossref","unstructured":"The Nguyen PN et al. Review of the epidemiology, diagnosis and management of invasive meningococcal disease in Vietnam. 2023;19(1):2172922.","DOI":"10.1080\/21645515.2023.2172922"},{"key":"688_CR5","doi-asserted-by":"crossref","unstructured":"Van CP et al. Invasive meningococcal disease remains a health threat in Vietnam People\u2019s Army. 2021:5261\u20135269.","DOI":"10.2147\/IDR.S339110"},{"key":"688_CR6","unstructured":"Kien NX. and N.V.B, Epidemiological features, common serogroups and risk factors for meningococcal infection. Vietnam Med J. 2023;526(1A)."},{"key":"688_CR7","doi-asserted-by":"crossref","unstructured":"V\u00e1zquez J et al. Global meningococcal initiative: guidelines for diagnosis and confirmation of invasive meningococcal disease. 2016;144(14):3052\u20137.","DOI":"10.1017\/S0950268816001308"},{"key":"688_CR8","doi-asserted-by":"crossref","unstructured":"Diallo K et al. Molecular diagnostic assays for the detection of common bacterial meningitis pathogens: A narrative review. 2021;65.","DOI":"10.1016\/j.ebiom.2021.103274"},{"issue":"5","key":"688_CR9","first-page":"712","volume":"81","author":"AR Feagins","year":"2020","unstructured":"Feagins AR, et al. Next generation rapid diagn tests meningitis diagnosis. 2020;81(5):712\u20138.","journal-title":"Next Generation Rapid Diagn Tests Meningitis Diagnosis"},{"key":"688_CR10","doi-asserted-by":"crossref","unstructured":"Trung NT et al. CRISPR-Cas12a combination to alleviate the false-positive in loop-mediated isothermal amplification-based diagnosis of Neisseria meningitidis. 2022;22(1):1\u20135.","DOI":"10.1186\/s12879-022-07363-w"},{"key":"688_CR11","doi-asserted-by":"crossref","unstructured":"Suleman E, Mtshali MS, J.J.o.V E. Investigation of false positives associated with loop-mediated isothermal amplification assays for detection of Toxoplasma Gondii in archived tissue samples of captive felids. 2016;28(5):536\u201342.","DOI":"10.1177\/1040638716659864"},{"issue":"1707","key":"688_CR12","first-page":"20150496","volume":"371","author":"F Hille","year":"2016","unstructured":"Hille F and E.J.P., Charpentier. CRISPR-Cas: biology mech relevance. 2016;371(1707):20150496.","journal-title":"CRISPR-Cas: Biology Mech Relevance"},{"key":"688_CR13","doi-asserted-by":"crossref","unstructured":"Li S-Y et al. CRISPR-Cas12a has both cis-and trans-cleavage activities on single-stranded DNA. 2018;28(4):491\u20133.","DOI":"10.1038\/s41422-018-0022-x"},{"key":"688_CR14","doi-asserted-by":"crossref","unstructured":"Broughton JP et al. CRISPR\u2013Cas12-based detection of SARS-CoV-2. 2020;38(7):870\u2013874.","DOI":"10.1038\/s41587-020-0513-4"},{"key":"688_CR15","unstructured":"Manual, editor. 2023, ed. C. 1.1.6. 2023: Paris: World Organisation for Animal Health."},{"key":"688_CR16","unstructured":"\u00c1lvarez SI, F.A.B.J. M.A.t.q.c. Andreu, Procedures for validation of diagnostic methods in clinical laboratory accredited by ISO 15189. 2011:209\u201329."},{"key":"688_CR17","doi-asserted-by":"crossref","unstructured":"Nutz S et al. Determination of the LOQ in real-time PCR by receiver operating characteristic curve analysis: application to qPCR assays for Fusarium verticillioides and F. Proliferatum. 2011;401:717\u201326.","DOI":"10.1007\/s00216-011-5089-x"},{"key":"688_CR18","doi-asserted-by":"crossref","unstructured":"Kralik P. and M.J.F.i.m. Ricchi. A basic guide to real time PCR in microbial diagnostics: definitions, parameters, and everything. 2017;8:108.","DOI":"10.3389\/fmicb.2017.00108"},{"key":"688_CR19","doi-asserted-by":"crossref","unstructured":"Frosch M et al. Conserved outer membrane protein of Neisseria meningitidis involved in capsule expression. 1992;60(3):798\u2013803.","DOI":"10.1128\/iai.60.3.798-803.1992"},{"key":"688_CR20","doi-asserted-by":"crossref","unstructured":"Lee D et al. Clinical evaluation of a loop-mediated isothermal amplification (LAMP) assay for rapid detection of Neisseria meningitidis in cerebrospinal fluid. 2015;10(4):e0122922.","DOI":"10.1371\/journal.pone.0122922"},{"key":"688_CR21","doi-asserted-by":"crossref","unstructured":"Mabey D et al. Diagnostics for the developing world. 2004;2(3):231\u2013240.","DOI":"10.1038\/nrmicro841"}],"container-title":["Annals of Clinical Microbiology and Antimicrobials"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1186\/s12941-024-00688-1.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/article\/10.1186\/s12941-024-00688-1\/fulltext.html","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1186\/s12941-024-00688-1.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,3,30]],"date-time":"2024-03-30T18:02:03Z","timestamp":1711821723000},"score":1,"resource":{"primary":{"URL":"https:\/\/ann-clinmicrob.biomedcentral.com\/articles\/10.1186\/s12941-024-00688-1"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,3,30]]},"references-count":21,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2024,12]]}},"alternative-id":["688"],"URL":"https:\/\/doi.org\/10.1186\/s12941-024-00688-1","relation":{},"ISSN":["1476-0711"],"issn-type":[{"value":"1476-0711","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,3,30]]},"assertion":[{"value":"2 February 2024","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"21 March 2024","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"30 March 2024","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Declarations"}},{"value":"All samples used in this study were obtained from patients admitted to 108 Military Central Hospital (108-MCH), Hanoi, Vietnam. The study was approved by the institutional review board and an independent ethics committee of 108 Military Central Hospital Hanoi, Vietnam (108MCH-# 2723\/H\u0110\u0110\u0110- 25062020). Written informed consent was obtained from all study patients and\/or their guardians. All samples used in this study were pseudo-anonymized prior to laboratory testing to de-identify the patients.","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Ethics approval and consent to participate"}},{"value":"All authors read and consented to publish the article.","order":3,"name":"Ethics","group":{"name":"EthicsHeading","label":"Consent for publication"}},{"value":"The authors declare no competing interests.","order":4,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"28"}}