{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,4]],"date-time":"2026-06-04T10:17:41Z","timestamp":1780568261859,"version":"3.54.1"},"reference-count":40,"publisher":"F1000 Research Ltd","license":[{"start":{"date-parts":[[2020,7,29]],"date-time":"2020-07-29T00:00:00Z","timestamp":1595980800000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"DELTAS Africa Initiative","award":["107742\/Z\/15\/Z"],"award-info":[{"award-number":["107742\/Z\/15\/Z"]}]},{"name":"Medical Research Council","award":["MR\/P024548\/1"],"award-info":[{"award-number":["MR\/P024548\/1"]}]}],"content-domain":{"domain":["aasopenresearch.org"],"crossmark-restriction":false},"short-container-title":["AAS Open Res"],"abstract":"<ns5:p>\n                    <ns5:bold>Background<\/ns5:bold>\n                    : Tuberculin skin test and interferon gamma release assay (IGRA) show limitations in diagnosing latent tuberculosis infection (LTBI) and poorly predict progression to active tuberculosis. This study will explore detection of\n                    <ns5:italic>Mycobacterium tuberculosis<\/ns5:italic>\n                    (\n                    <ns5:italic>M.tb<\/ns5:italic>\n                    ) DNA in CD34\n                    <ns5:sup>+<\/ns5:sup>\n                    peripheral blood mononuclear cells (PBMCs) as a biomarker for LTBI and monitoring chemoprophylaxis response.\n                  <\/ns5:p>\n                  <ns5:p>\n                    <ns5:bold>Methods:<\/ns5:bold>\n                    In a cross-sectional study, 120 household contacts (60 HIV positive and 60 HIV negative) will be recruited. Also, 10 patients with sputum positive pulmonary tuberculosis and 10 visitors from low incidence countries with no history of TB treatment will be recruited as positive and negative controls, respectively. Participants will donate 100 ml (50 ml for TB patients) of blood to isolate PBMCs using density gradient centrifugation. Isolated PBMCs will be separated into CD34\n                    <ns5:sup>+<\/ns5:sup>\n                    and CD34\n                    <ns5:sup>-<\/ns5:sup>\n                    enriched cellular fractions. DNA from each fraction will be purified, quantified and subjected to droplet digital PCR targeting\n                    <ns5:italic>IS6110<\/ns5:italic>\n                    (a\n                    <ns5:italic>M.tb<\/ns5:italic>\n                    Complex multi-copy gene) and\n                    <ns5:italic>rpoB<\/ns5:italic>\n                    , a single copy gene. Also, 4 ml of blood will be drawn for IGRA. In a nested prospective study, 60 HIV positive participants will be given 300 mg of Isoniazid Preventive Therapy (IPT) daily for six months, after which they will donate a second 100 ml blood sample that will be processed as described above.\n                  <\/ns5:p>\n                  <ns5:p>\n                    Data from the cross-sectional study will be analysed to determine the proportion of individuals in whom\n                    <ns5:italic>M.tb<\/ns5:italic>\n                    DNA is detectable in CD34\n                    <ns5:sup>+<\/ns5:sup>\n                    and CD34\n                    <ns5:sup>-<\/ns5:sup>\n                    fractions and number of\n                    <ns5:italic>M.tb<\/ns5:italic>\n                    genomes present. Data from the prospective study will be analysed to compare the proportion of individuals with detectable\n                    <ns5:italic>M.tb<\/ns5:italic>\n                    DNA in CD34\n                    <ns5:sup>+<\/ns5:sup>\n                    and CD34\n                    <ns5:sup>-<\/ns5:sup>\n                    fractions, and median\n                    <ns5:italic>M.tb<\/ns5:italic>\n                    genome copy number, post vs pre-IPT.\n                  <\/ns5:p>\n                  <ns5:p>\n                    <ns5:bold>Discussion:<\/ns5:bold>\n                    This study will determine whether detection of\n                    <ns5:italic>M.tb<\/ns5:italic>\n                    DNA in CD34\n                    <ns5:sup>+<\/ns5:sup>\n                    PBMCs holds promise as a biomarker for LTBI and monitoring chemoprophylaxis response.\n                  <\/ns5:p>","DOI":"10.12688\/aasopenres.13108.1","type":"journal-article","created":{"date-parts":[[2020,7,29]],"date-time":"2020-07-29T06:20:26Z","timestamp":1596003626000},"page":"34","update-policy":"https:\/\/doi.org\/10.12688\/aasopenres.crossmark-policy","source":"Crossref","is-referenced-by-count":4,"title":["Detection of Mycobacterium tuberculosis DNA in CD34+ peripheral blood mononuclear cells of Ugandan adults with latent infection: a cross-sectional and nested prospective study"],"prefix":"10.12688","volume":"3","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-9611-2178","authenticated-orcid":false,"given":"Jonathan","family":"Mayito","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8303-6046","authenticated-orcid":false,"given":"Irene","family":"Andia Biraro","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stephen","family":"T. Reece","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Adrian","family":"R. Martineau","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8075-3069","authenticated-orcid":false,"given":"David","family":"P. Kateete","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"2560","published-online":{"date-parts":[[2020,7,29]]},"reference":[{"key":"ref-1","article-title":"Global Tuberculosis Report 2017","year":"2017"},{"key":"ref-2","doi-asserted-by":"publisher","first-page":"2","DOI":"10.3389\/fmicb.2011.00002","article-title":"Mycobacterium tuberculosis Infection and Inflammation: what is Beneficial for the Host and for the Bacterium?","volume":"2","author":"S Sasindran","year":"2011","journal-title":"Front Microbiol."},{"key":"ref-3","doi-asserted-by":"publisher","first-page":"36-42","DOI":"10.1164\/rccm.2109089","article-title":"Transmission dynamics of tuberculosis in Tarrant county, Texas.","volume":"166","author":"S Weis","year":"2002","journal-title":"Am J Respir Crit Care Med."},{"key":"ref-4","doi-asserted-by":"publisher","first-page":"1703-9","DOI":"10.1056\/NEJM199406163302402","article-title":"The epidemiology of tuberculosis in San Francisco. A population-based study using conventional and molecular methods.","volume":"330","author":"P Small","year":"1994","journal-title":"N Engl J Med."},{"key":"ref-5","doi-asserted-by":"publisher","first-page":"1009-21","DOI":"10.1001\/archinte.163.9.1009","article-title":"The growing burden of tuberculosis: global trends and interactions with the HIV epidemic.","volume":"163","author":"E Corbett","year":"2003","journal-title":"Arch Intern Med."},{"key":"ref-6","doi-asserted-by":"publisher","first-page":"e1002152","DOI":"10.1371\/journal.pmed.1002152","article-title":"The Global Burden of Latent Tuberculosis Infection: A Re-estimation Using Mathematical Modelling.","volume":"13","author":"R Houben","year":"2016","journal-title":"PLoS Med."},{"key":"ref-7","doi-asserted-by":"publisher","first-page":"131-8","DOI":"10.1093\/oxfordjournals.aje.a121593","article-title":"The prognosis of a positive tuberculin reaction in childhood and adolescence.","volume":"99","author":"G Comstock","year":"1974","journal-title":"Am J Epidemiol."},{"key":"ref-8","doi-asserted-by":"publisher","first-page":"784-91","DOI":"10.1093\/cid\/cir951","article-title":"Risk of progression to active tuberculosis following reinfection with Mycobacterium tuberculosis.","volume":"54","author":"J Andrews","year":"2012","journal-title":"Clin Infect Dis."},{"key":"ref-9","article-title":"Global Tuberculosis Report","year":"2015"},{"key":"ref-10","doi-asserted-by":"publisher","first-page":"2127-2135","DOI":"10.1056\/NEJMra1405427","article-title":"Latent Mycobacterium tuberculosis Infection.","volume":"372","author":"G Haileyesus","year":"2015","journal-title":"N Engl J Med."},{"key":"ref-11","doi-asserted-by":"publisher","first-page":"729-736","DOI":"10.1016\/j.jmii.2014.08.010","article-title":"Prevalence of latent tuberculosis infection in persons with and without human immunodeficiency virus infection using two interferon-gamma release assays and tuberculin skin test in a low human immunodeficiency virus prevalence, intermediate tuberculosis-burden country.","volume":"49","author":"W Lin","year":"2016","journal-title":"J Microbiol Immunol Infect."},{"key":"ref-12","doi-asserted-by":"publisher","first-page":"63-72","DOI":"10.2147\/idr.s8994","article-title":"The use of anti-tuberculosis therapy for latent TB infection.","volume":"3","author":"J Denholm","year":"2010","journal-title":"Infect Drug Resist."},{"key":"ref-13","doi-asserted-by":"publisher","first-page":"199-207","DOI":"10.1007\/s13665-013-0064-y","article-title":"Diagnosis and treatment of latent tuberculosis infection: an update.","volume":"2","author":"A Person","year":"2013","journal-title":"Curr Respir Care Rep."},{"key":"ref-14","doi-asserted-by":"publisher","first-page":"619","DOI":"10.1186\/s12879-019-4238-2","article-title":"The association between tuberculin skin test result and active tuberculosis risk of college students in Beijing, China: a retrospective cohort study.","volume":"19","author":"D Cao","year":"2019","journal-title":"BMC Infect Dis."},{"key":"ref-15","doi-asserted-by":"publisher","first-page":"150-3","DOI":"10.1097\/INF.0b013e318236ae2b","article-title":"A strong tuberculin reaction in primary school children predicts tuberculosis in adolescence.","volume":"31","author":"C Leung","year":"2012","journal-title":"Pediatr Infect Dis J."},{"key":"ref-16","doi-asserted-by":"publisher","first-page":"968-75","DOI":"10.1093\/clinids\/17.6.968","article-title":"The tuberculin skin test.","volume":"17","author":"R Huebner","year":"1993","journal-title":"Clin Infect Dis."},{"key":"ref-17","first-page":"222-7","article-title":"Effect of non-tuberculous Mycobacteria infection on tuberculin results among primary school children in Kenya.","volume":"72","author":"D Kwamanga","year":"1995","journal-title":"East Afr Med J."},{"key":"ref-18","doi-asserted-by":"publisher","first-page":"63-75","DOI":"10.1378\/chest.11-3157","article-title":"Predictive value of interferon-\u03b3 release assays and tuberculin skin testing for progression from latent TB infection to disease state: a meta-analysis.","volume":"142","author":"R Diel","year":"2012","journal-title":"Chest."},{"key":"ref-19","doi-asserted-by":"publisher","first-page":"804-9","DOI":"10.1136\/thorax.57.9.804","article-title":"A meta-analysis of the effect of Bacille Calmette Guerin vaccination on tuberculin skin test measurements.","volume":"57","author":"L Wang","year":"2002","journal-title":"Thorax."},{"key":"ref-20","doi-asserted-by":"publisher","first-page":"814943","DOI":"10.1155\/2011\/814943","article-title":"Pathogenesis, immunology, and diagnosis of latent Mycobacterium tuberculosis infection.","volume":"2011","author":"S Ahmad","year":"2011","journal-title":"Clin Dev Immunol."},{"key":"ref-21","doi-asserted-by":"publisher","first-page":"177-184","DOI":"10.7326\/0003-4819-149-3-200808050-00241","article-title":"Systematic review: T-cell-based assays for the diagnosis of latent tuberculosis infection: an update.","volume":"149","author":"M Pai","year":"2008","journal-title":"Ann Intern Med."},{"key":"ref-22","doi-asserted-by":"publisher","first-page":"271-284","DOI":"10.1097\/MCP.0b013e328338094f","article-title":"New and improved tuberculosis diagnostics: evidence, policy, practice, and impact.","volume":"16","author":"M Pai","year":"2010","journal-title":"Curr Opin Pulm Med."},{"key":"ref-23","doi-asserted-by":"publisher","first-page":"CD001363","DOI":"10.1002\/14651858.CD001363","article-title":"Isoniazid for preventing tuberculosis in non-HIV infected persons.","volume":"1999","author":"M Smieja","year":"2000","journal-title":"Cochrane Database Syst Rev."},{"key":"ref-24","doi-asserted-by":"publisher","first-page":"216-9","DOI":"10.1038\/nature10160","article-title":"In vivo imaging of Treg cells providing immune privilege to the haematopoietic stem-cell niche.","volume":"474","author":"J Fujisaki","year":"2011","journal-title":"Nature."},{"key":"ref-25","doi-asserted-by":"publisher","first-page":"380-90","DOI":"10.1016\/j.stem.2010.07.011","article-title":"The distinct metabolic profile of hematopoietic stem cells reflects their location in a hypoxic niche.","volume":"7","author":"T Simsek","year":"2010","journal-title":"Cell Stem Cell."},{"key":"ref-26","doi-asserted-by":"publisher","first-page":"5431-5436","DOI":"10.1073\/pnas.0701152104","article-title":"Distribution of hematopoietic stem cells in the bone marrow according to regional hypoxia.","volume":"104","author":"K Parmar","year":"2007","journal-title":"Proc Natl Acad Sci USA."},{"key":"ref-27","doi-asserted-by":"publisher","first-page":"1797-1806","DOI":"10.1084\/jem.183.4.1797","article-title":"Isolation and functional properties of murine hematopoietic stem cells that are replicating In vivo.","volume":"183","author":"M Goodell","year":"1996","journal-title":"J Exp Med."},{"key":"ref-28","doi-asserted-by":"publisher","first-page":"1028-1034","DOI":"10.1038\/nm0901-1028","article-title":"The ABC transporter Bcrp1\/ABCG2 is expressed in a wide variety of stem cells and is a molecular determinant of the side-population phenotype.","volume":"7","author":"S Zhou","year":"2001","journal-title":"Nat Med."},{"key":"ref-29","doi-asserted-by":"publisher","first-page":"21653-8","DOI":"10.1073\/pnas.1007967107","article-title":"Mycobacterium tuberculosis evades host immunity by recruiting mesenchymal stem cells.","volume":"107","author":"S Raghuvanshi","year":"2010","journal-title":"Proc Natl Acad Sci U S A."},{"key":"ref-30","doi-asserted-by":"publisher","first-page":"170ra13","DOI":"10.1126\/scitranslmed.3004912","article-title":"CD271+ bone marrow mesenchymal stem cells may provide a niche for dormant Mycobacterium tuberculosis.","volume":"5","author":"B Das","year":"2013","journal-title":"Sci Transl Med."},{"key":"ref-31","doi-asserted-by":"publisher","first-page":"e0169119","DOI":"10.1371\/journal.pone.0169119","article-title":"Human and Mouse Hematopoietic Stem Cells Are a Depot for Dormant Mycobacterium tuberculosis.","volume":"12","author":"J Tornack","year":"2017","journal-title":"PLoS One."},{"key":"ref-32","doi-asserted-by":"publisher","first-page":"1667-1671","DOI":"10.1093\/infdis\/jiy041","article-title":"Mycobacterium tuberculosis-Infected Hematopoietic Stem and Progenitor Cells Unable to Express Inducible Nitric Oxide Synthase Propagate Tuberculosis in Mice.","volume":"217","author":"S Reece","year":"2018","journal-title":"J Infect Dis."},{"key":"ref-33","doi-asserted-by":"publisher","first-page":"161-78","DOI":"10.1016\/j.it.2015.01.003","article-title":"Nitric oxide synthase in innate and adaptive immunity: an update.","volume":"36","author":"C Bogdan","year":"2015","journal-title":"Trends Immunol."},{"key":"ref-34","doi-asserted-by":"crossref","DOI":"10.12688\/aasopenres.13108.1","article-title":"Detection of Mycobacterium tuberculosis DNA in CD34+ peripheral blood mononuclear cells of Ugandan adults with latent infection: A cross-sectional & nested prospective study","author":"J Mayito","year":"2020"},{"key":"ref-35","doi-asserted-by":"publisher","first-page":"3-4","DOI":"10.1002\/9780471462422.eoct005","article-title":"Sample Size Calculation for Comparing Proportions.","author":"H Wang","year":"2007","journal-title":"Wiley Encyclopedia of Clinical Trials."},{"key":"ref-36","first-page":"232","article-title":"Fundamentals of Biostatistics.","author":"B Rosner","year":"1995"},{"key":"ref-37","doi-asserted-by":"publisher","first-page":"41933","DOI":"10.1038\/srep41933","article-title":"Association of Body Mass Index with the Tuberculosis Infection: a Population-based Study among 17796 Adults in Rural China.","volume":"7","author":"H Zhang","year":"2017","journal-title":"Sci Rep."},{"key":"ref-38","doi-asserted-by":"publisher","first-page":"E301-e303","DOI":"10.21037\/jtd.2017.03.47","article-title":"The link between tuberculosis and body mass index.","volume":"9","author":"A Casha","year":"2017","journal-title":"J Thorac Dis."},{"key":"ref-39","doi-asserted-by":"publisher","first-page":"e016961","DOI":"10.1136\/bmjopen-2017-016961","article-title":"Incidence and determinants of tuberculosis infection among adult patients with HIV attending HIV care in north-east Ethiopia: a retrospective cohort study.","volume":"8","author":"A Ahmed","year":"2018","journal-title":"BMJ Open."},{"key":"ref-40","doi-asserted-by":"publisher","first-page":"165-203","DOI":"10.1385\/1-59259-147-7:165","article-title":"Restriction fragment length polymorphism typing of mycobacteria.","volume":"54","author":"D van Soolingen","year":"2001","journal-title":"Methods Mol Med."}],"container-title":["AAS Open Research"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/aasopenresearch.org\/articles\/3-34\/v1\/xml","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/aasopenresearch.org\/articles\/3-34\/v1\/pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/aasopenresearch.org\/articles\/3-34\/v1\/iparadigms","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2020,8,13]],"date-time":"2020-08-13T04:45:54Z","timestamp":1597293954000},"score":1,"resource":{"primary":{"URL":"https:\/\/aasopenresearch.org\/articles\/3-34\/v1"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,7,29]]},"references-count":40,"URL":"https:\/\/doi.org\/10.12688\/aasopenres.13108.1","relation":{"has-review":[{"id-type":"doi","id":"10.21956\/aasopenres.14208.r27716","asserted-by":"subject"},{"id-type":"doi","id":"10.21956\/aasopenres.14208.r27717","asserted-by":"subject"},{"id-type":"doi","id":"10.21956\/aasopenres.14208.r27716","asserted-by":"object"},{"id-type":"doi","id":"10.21956\/aasopenres.14208.r27717","asserted-by":"object"}]},"ISSN":["2515-9321"],"issn-type":[{"value":"2515-9321","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,7,29]]},"assertion":[{"value":"Indexed","URL":"https:\/\/aasopenresearch.org\/articles\/3-34\/v1#article-reports","order":0,"name":"referee-status","label":"Referee status","group":{"name":"current-referee-status","label":"Current Referee Status"}},{"value":"10.21956\/aasopenres.14208.r27716, Emmanuel O. Babafemi, School of Pharmacy and Biomolecular Sciences, Liverpool John Moores University, Liverpool, UK, 10 Aug 2020, version 1, indexed","URL":"https:\/\/aasopenresearch.org\/articles\/3-34\/v1#referee-response-27716","order":0,"name":"referee-response-27716","label":"Referee Report","group":{"name":"article-reports","label":"Article Reports"}},{"value":"<b>Jonathan Mayito<\/b>; \n<i>Posted: 10 Aug 2020<\/i>; Thank you Emmanuel, for your time to review this article and the positive comments. Thank you too for having the same enthusiasm about this work that seeks to bridge the gap in the diagnosis of latent tuberculosis through evaluating CD34+PBMCs as target cells for detecting M.tb during latency.","URL":"https:\/\/aasopenresearch.org\/articles\/3-34\/v1#referee-comment-3160","order":1,"name":"referee-comment-3160","label":"Referee Comment","group":{"name":"article-reports","label":"Article Reports"}},{"value":"10.21956\/aasopenres.14208.r27717, Masaki Yamamoto, Respiratory Disease Center, Yokohama City University Medical Center, Yokohama, Japan, 11 Aug 2020, version 1, indexed","URL":"https:\/\/aasopenresearch.org\/articles\/3-34\/v1#referee-response-27717","order":2,"name":"referee-response-27717","label":"Referee Report","group":{"name":"article-reports","label":"Article Reports"}},{"value":"<b>Jonathan Mayito<\/b>; \n<i>Posted: 12 Aug 2020<\/i>; Thank you, Masaki for your time to review our article. I agree with you that a comparative prospective design including HIV positive participants not on IPT would have been more appropriate to delineate the effect of IPT. However, as you point out this would have been unethical given that IPT is standard of care. Interesting suggestion to compare the adherent and the poorly adherent. We will look into this at analysis.","URL":"https:\/\/aasopenresearch.org\/articles\/3-34\/v1#referee-comment-3161","order":3,"name":"referee-comment-3161","label":"Referee Comment","group":{"name":"article-reports","label":"Article Reports"}},{"value":"This work is funded by grants from the Medical Research Council (ref. MR\/P024548\/1; to A. R. M.) and the DELTAS Africa Initiative (ref. DEL-15-011 [to J. M.], via THRiVE-2). The DELTAS Africa Initiative is an independent funding scheme of the AAS\u2019s Alliance for Accelerating Excellence in Science in Africa and supported by the NEPAD Agency with funding from the Wellcome Trust Grant No. 107742\/Z\/15\/Z and the United Kingdom government. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.","order":4,"name":"grant-information","label":"Grant Information"},{"value":"This is an open access article distributed under the terms of the Creative Commons Attribution Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.","order":0,"name":"copyright-info","label":"Copyright"}]}}