{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,19]],"date-time":"2026-08-19T20:04:24Z","timestamp":1787169864092,"version":"build-2736575974"},"reference-count":25,"publisher":"Oxford University Press (OUP)","issue":"4","license":[{"start":{"date-parts":[[2024,3,14]],"date-time":"2024-03-14T00:00:00Z","timestamp":1710374400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2024,4,3]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:sec>\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>Improved monitoring of Mycobacterium tuberculosis response to treatment is urgently required. We previously developed the molecular bacterial load assay (MBLA), but it is challenging to integrate into the clinical diagnostic laboratory due to a labor-intensive protocol required at biosafety level 3 (BSL-3). A modified assay was needed.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>The rapid enumeration and diagnostic for tuberculosis (READ-TB) assay was developed. Acetic acid was tested and compared to 4\u2005M guanidine thiocyanate to be simultaneously bactericidal and preserve mycobacterial RNA. The extraction was based on silica column technology and incorporated low-cost reagents: 3\u2005M sodium acetate and ethanol for the RNA extraction to replace phenol\u2013chloroform. READ-TB was fully validated and compared directly to the MBLA using sputa collected from individuals with tuberculosis.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>Acetic acid was bactericidal to M. tuberculosis with no significant loss in 16S rRNA or an unprotected mRNA fragment when sputum was stored in acetic acid at 25\u00b0C for 2 weeks or \u221220\u00b0C for 1 year. This novel use of acetic acid allows processing of sputum for READ-TB at biosafety level 2 (BSL-2) on sample receipt. READ-TB is semiautomated and rapid. READ-TB correlated with the MBLA when 85 human sputum samples were directly compared (R2 = 0.74).<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>READ-TB is an improved version of the MBLA and is available to be adopted by clinical microbiology laboratories as a tool for tuberculosis treatment monitoring. READ-TB will have a particular impact in low- and middle-income countries (LMICs) for laboratories with no BSL-3 laboratory and for clinical trials testing new combinations of anti-tuberculosis drugs.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1093\/clinchem\/hvae013","type":"journal-article","created":{"date-parts":[[2024,1,18]],"date-time":"2024-01-18T15:14:20Z","timestamp":1705590860000},"page":"642-652","source":"Crossref","is-referenced-by-count":5,"title":["Acetic Acid Enables Molecular Enumeration of\n                    <i>Mycobacterium tuberculosis<\/i>\n                    from Sputum and Eliminates the Need for a Biosafety Level 3 Laboratory"],"prefix":"10.1093","volume":"70","author":[{"given":"Ana","family":"Palekyte","sequence":"first","affiliation":[{"name":"Centre for Clinical Microbiology, Division of Infection and Immunity, University College London , London , United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anna","family":"Morkowska","sequence":"additional","affiliation":[{"name":"Imperial College London, Infection and Immunity, Department of Microbiology, Charing Cross Hospital , London , United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Owen","family":"Billington","sequence":"additional","affiliation":[{"name":"Health Services Laboratories , London , United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6877-2531","authenticated-orcid":false,"given":"Stephen","family":"Morris-Jones","sequence":"additional","affiliation":[{"name":"Department of Clinical Microbiology, University College London Hospitals NHS Foundation Trust , London , United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"James","family":"Millard","sequence":"additional","affiliation":[{"name":"Liverpool Centre for Global Health Research, University of Liverpool , Liverpool , United Kingdom"},{"name":"Guy\u2019s and St Thomas\u2019 NHS Foundation Trust , London , United Kingdom"},{"name":"Africa Health Research Institute , Durban , South Africa"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mohlopheni J","family":"Marakalala","sequence":"additional","affiliation":[{"name":"Africa Health Research Institute , Durban , South Africa"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Olumuyiwa","family":"Owolabi","sequence":"additional","affiliation":[{"name":"Vaccines and Immunity, MRC Unit The Gambia at The London School of Hygiene and Tropical Medicine , Banjul , The Gambia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Basil","family":"Sambou","sequence":"additional","affiliation":[{"name":"Vaccines and Immunity, MRC Unit The Gambia at The London School of Hygiene and Tropical Medicine , Banjul , The Gambia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alimuddin","family":"Zumla","sequence":"additional","affiliation":[{"name":"Centre for Clinical Microbiology, Division of Infection and Immunity, University College London , London , United Kingdom"},{"name":"National Institute of Health and Research Biomedical Research Centre, UCL Hospitals National Health Service Foundation Trust , London , United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jayne S","family":"Sutherland","sequence":"additional","affiliation":[{"name":"Vaccines and Immunity, MRC Unit The Gambia at The London School of Hygiene and Tropical Medicine , Banjul , The Gambia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Timothy D","family":"McHugh","sequence":"additional","affiliation":[{"name":"Centre for Clinical Microbiology, Division of Infection and Immunity, University College London , London , United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0596-758X","authenticated-orcid":false,"given":"Isobella","family":"Honeyborne","sequence":"additional","affiliation":[{"name":"Centre for Clinical Microbiology, Division of Infection and Immunity, University College London , London , United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2024,3,14]]},"reference":[{"key":"2024040223561056500_hvae013-B1","volume-title":"Global tuberculosis report 2023","author":"WHO","year":"2023"},{"key":"2024040223561056500_hvae013-B2","volume-title":"WHO Consolidated guidelines on tuberculosis. 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