{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,11]],"date-time":"2026-04-11T20:52:47Z","timestamp":1775940767620,"version":"3.50.1"},"reference-count":47,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2019,12,13]],"date-time":"2019-12-13T00:00:00Z","timestamp":1576195200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Diagnostics"],"abstract":"<jats:p>Tuberculosis (TB) is the leading cause of death globally, surpassing HIV. Furthermore, multidrug-resistant and extensively drug-resistant TB have become global public health threats. Care of TB patients starts with quality, accessible, and affordable diagnosis. The study presents a novel technique called nanoparticle-based colorimetric biosensing assay (NCBA) based on the principles of magnetically activated cell enrichment. A total of 1108 sputum samples were subjected to sputum smear microscopy (SSM), NCBA, and standard culture. SSM and NCBA were completed in 20 min; culture was completed in 8 weeks. Results show that NCBA has matching sensitivity of 100.0% and specificity of 99.7% compared to the gold standard culture method at a cost of $0.50\/test based on Peruvian conditions. Sputum smear microscopy has 63.87% sensitivity compared to culture. NCBA has the potential of being used in local health clinics as it only requires a microscope that is widely available in many rural areas. Because NCBA could detect low levels of bacterial load comparable to culture, it could be used for rapid and early TB-onset detection. The gain in time is critical as TB is airborne and highly infectious, minimizing contact exposure. Early detection could lead to early treatment, while the patient\u2019s immune system is still high. The low cost makes NCBA affordable and accessible to those who need them the most.<\/jats:p>","DOI":"10.3390\/diagnostics9040222","type":"journal-article","created":{"date-parts":[[2019,12,13]],"date-time":"2019-12-13T11:27:22Z","timestamp":1576236442000},"page":"222","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":22,"title":["Nanoparticle-Based Biosensing Assay for Universally Accessible Low-Cost TB Detection with Comparable Sensitivity as Culture"],"prefix":"10.3390","volume":"9","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4360-2100","authenticated-orcid":false,"given":"Ruben Kenny","family":"Briceno","sequence":"first","affiliation":[{"name":"Instituto de Investigacion en Ciencia y Tecnologia, Universidad Cesar Vallejo, Trujillo 13100, Peru"},{"name":"Hospital Victor Lazarte Echegaray, Trujillo 13100, Peru"},{"name":"Institute for Global Health, Michigan State University, East Lansing, MI 48824, USA"},{"name":"Global Alliance for Rapid Diagnostics, Michigan State University, East Lansing, MI 48824, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shane Ryan","family":"Sergent","sequence":"additional","affiliation":[{"name":"Instituto de Investigacion en Ciencia y Tecnologia, Universidad Cesar Vallejo, Trujillo 13100, Peru"},{"name":"Institute for Global Health, Michigan State University, East Lansing, MI 48824, USA"},{"name":"Kingman Regional Medical Center, Kingman, AZ 86409, USA"},{"name":"Global Alliance for Rapid Diagnostics, Michigan State University, East Lansing, MI 48824, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Santiago Moises","family":"Benites","sequence":"additional","affiliation":[{"name":"Instituto de Investigacion en Ciencia y Tecnologia, Universidad Cesar Vallejo, Trujillo 13100, Peru"},{"name":"Global Alliance for Rapid Diagnostics, Michigan State University, East Lansing, MI 48824, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Evangelyn C.","family":"Alocilja","sequence":"additional","affiliation":[{"name":"Global Alliance for Rapid Diagnostics, Michigan State University, East Lansing, MI 48824, USA"},{"name":"Biosystems and Agricultural Engineering, Michigan State University, East Lansing, MI 48824, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2019,12,13]]},"reference":[{"key":"ref_1","unstructured":"(2019, December 11). TB statistics | Incidence, prevalence, high burden. Available online: https:\/\/tbfacts.org\/tb-statistics\/."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"314","DOI":"10.1128\/CMR.00059-10","article-title":"Laboratory Diagnosis of Tuberculosis in Resource-Poor Countries: Challenges and Opportunities","volume":"24","author":"Parsons","year":"2011","journal-title":"Clin. Microbiol. Rev."},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Yang, T., Zhong, J., Zhang, J., Li, C., Yu, X., Xiao, J., Jia, X., Ding, N., Ma, G., and Wang, G. (2018). Pan-Genomic Study of Mycobacterium tuberculosis Reflecting the Primary\/Secondary Genes, Generality\/Individuality, and the Interconversion Through Copy Number Variations. Front. Microbiol., 9.","DOI":"10.3389\/fmicb.2018.01886"},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Sakamoto, H., Lee, S., Ishizuka, A., Hinoshita, E., Hori, H., Ishibashi, N., Komada, K., Norizuki, M., Katsuma, Y., and Akashi, H. (2019). Challenges and opportunities for eliminating tuberculosis \u2013 leveraging political momentum of the UN high-level meeting on tuberculosis. BMC Public Health, 19.","DOI":"10.1186\/s12889-019-6399-8"},{"key":"ref_5","unstructured":"(2019, September 18). Tuberculosis in Developing Countries, Available online: https:\/\/www.cdc.gov\/mmwr\/preview\/mmwrhtml\/00001729.htm."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.tube.2019.01.001","article-title":"Mycobacteria and their sweet proteins: An overview of protein glycosylation and lipoglycosylation in M. tuberculosis","volume":"115","author":"Mehaffy","year":"2019","journal-title":"Tuberculosis"},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"MacNeil, A. (2019). Global Epidemiology of Tuberculosis and Progress Toward Achieving Global Targets \u2014 2017. MMWR Morb. Mortal. Wkly. Rep., 68.","DOI":"10.15585\/mmwr.mm6811a3"},{"key":"ref_8","unstructured":"(2018, May 01). Issues and threats of Tuberculosis in Nepal | Connecting our members with evidence and expertise. Available online: https:\/\/www.ghdonline.org\/ic\/discussion\/issues-and-threats-of-tuberculosis-in-nepal\/."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"94","DOI":"10.1128\/AAC.4.2.94","article-title":"Enumeration of Tubercle Bacilli in Sputum of Patients with Pulmonary Tuberculosis","volume":"4","author":"Hobby","year":"1973","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"664","DOI":"10.1016\/S1473-3099(06)70602-8","article-title":"Sputum processing methods to improve the sensitivity of smear microscopy for tuberculosis: a systematic review","volume":"6","author":"Steingart","year":"2006","journal-title":"Lancet Infect. Dis."},{"key":"ref_11","first-page":"442","article-title":"Sputum smear microscopy in tuberculosis: Is it still relevant?","volume":"137","author":"Desikan","year":"2013","journal-title":"Indian J. Med. Res."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Olaru, I.D., Heyckendorf, J., Grossmann, S., and Lange, C. (2014). Time to Culture Positivity and Sputum Smear Microscopy during Tuberculosis Therapy. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0106075"},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Demers, A.-M., Verver, S., Boulle, A., Warren, R., van Helden, P., Behr, M.A., and Coetzee, D. (2012). High yield of culture-based diagnosis in a TB-endemic setting. BMC Infect. Dis., 12.","DOI":"10.1186\/1471-2334-12-218"},{"key":"ref_14","first-page":"435","article-title":"Comparison of culture, microscopic smear and molecular methods in diagnosis of tuberculosis","volume":"31","author":"Afsar","year":"2018","journal-title":"Rev. Esp. Quimioter."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1793","DOI":"10.1183\/09031936.00217313","article-title":"Replacing smear microscopy for the diagnosis of tuberculosis: what is the market potential?","volume":"43","author":"Kik","year":"2014","journal-title":"Eur. Respir. J."},{"key":"ref_16","first-page":"14","article-title":"Comparison of detection of Acid Fast Bacilli in clinical samples by AFB Smear microscopy and Culture in the diagnosis of Tuberculosis in a tertiary care setting","volume":"15","author":"Shabbir","year":"2009","journal-title":"Pak J Che Med"},{"key":"ref_17","unstructured":"Das, S., Narang, P., Nagamiah, S., Mishra, P.S., Deotale, V., and Mendiratta, D.K. (2019, September 30). Evaluation of variants of carbol fuchsin solution to stain acid-fast bacilli in-situ by the pot method. Available online: https:\/\/www.ingentaconnect.com\/content\/iuatld\/ijtld\/2015\/00000019\/00000012\/art00012%3bjsessionid=cb82amdbsq0bt.x-ic-live-03#."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1553","DOI":"10.1128\/JCM.00348-09","article-title":"High Sensitivity and Specificity of Acid-Fast Microscopy for Diagnosis of Pulmonary Tuberculosis in an African Population with a High Prevalence of Human Immunodeficiency Virus","volume":"47","author":"Shea","year":"2009","journal-title":"J. Clin. Microbiol."},{"key":"ref_19","first-page":"120","article-title":"Sputum processing prior to Mycobacterium tuberculosis detection by culture or nucleic acid amplification testing: a narrative review","volume":"5","author":"Allen","year":"2016","journal-title":"Res. Rev. J. Microbiol. Biotechnol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"537","DOI":"10.5588\/ijtld.14.0798","article-title":"The impact of sputum quality on tuberculosis diagnosis: a systematic review","volume":"19","author":"Ho","year":"2015","journal-title":"Int. J. Tuberc. Lung Dis."},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Nathavitharana, R.R., Cudahy, P.G.T., Schumacher, S.G., Steingart, K.R., Pai, M., and Denkinger, C.M. (2017). Accuracy of line probe assays for the diagnosis of pulmonary and multidrug-resistant tuberculosis: a systematic review and meta-analysis. Eur. Respir. J., 49.","DOI":"10.1183\/13993003.01075-2016"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1846","DOI":"10.1128\/JCM.03005-13","article-title":"Comparison of Xpert MTB\/RIF with Line Probe Assay for Detection of Rifampin-Monoresistant Mycobacterium tuberculosis","volume":"52","author":"Rufai","year":"2014","journal-title":"J. Clin. Microbiol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"70","DOI":"10.4103\/ijmr.IJMR_831_14","article-title":"Line probe assay for detection of Mycobacterium tuberculosis complex: An experience from Central India","volume":"145","author":"Desikan","year":"2017","journal-title":"Indian J. Med. Res."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1202","DOI":"10.1128\/JCM.02268-10","article-title":"Rapid Molecular Detection of Extrapulmonary Tuberculosis by the Automated GeneXpert MTB\/RIF System","volume":"49","author":"Hillemann","year":"2011","journal-title":"J. Clin. Microbiol."},{"key":"ref_25","unstructured":"(2019, December 06). WHO | Xpert MTB\/RIF - rapid TB test - WHO publishes policy and guidance for implementers. Available online: https:\/\/www.who.int\/tb\/features_archive\/xpert_rapid_tb_test\/en\/."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"e000132","DOI":"10.1136\/bmjgh-2016-000132","article-title":"Addressing the challenges of diagnostics demand and supply: insights from an online global health discussion platform","volume":"1","author":"Engel","year":"2016","journal-title":"BMJ Glob. Health"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"708","DOI":"10.1183\/09031936.00147912","article-title":"Xpert MTB\/RIF for diagnosis of tuberculosis and drug-resistant tuberculosis: a cost and affordability analysis","volume":"42","author":"Pantoja","year":"2013","journal-title":"Eur. Respir. J."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1763","DOI":"10.1183\/09031936.00193413","article-title":"Financial burden for tuberculosis patients in low- and middle-income countries: a systematic review","volume":"43","author":"Tanimura","year":"2014","journal-title":"Eur. Respir. J."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1038\/s41598-017-03930-3","article-title":"XtracTB Assay, a Mycobacterium tuberculosis molecular screening test with sensitivity approaching culture","volume":"7","author":"Reed","year":"2017","journal-title":"Sci. Rep."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"723","DOI":"10.5588\/ijtld.17.0835","article-title":"Global tuberculosis targets and milestones set for 2016\u20132035: definition and rationale","volume":"22","author":"Floyd","year":"2018","journal-title":"Int. J. Tuberc. Lung Dis."},{"key":"ref_31","unstructured":"(2019, October 23). Publicaciones de An\u00e1lisis de Situaci\u00f3n de Salud. Available online: https:\/\/www.dge.gob.pe\/portal\/index.php?option=com_content&view=article&id=255%20&Itemid=105."},{"key":"ref_32","unstructured":"(2019, October 23). WHO | Global tuberculosis report 2019. Available online: http:\/\/www.who.int\/tb\/publications\/global_report\/en\/."},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Gordillo-Marroqu\u00edn, C., G\u00f3mez-Velasco, A., S\u00e1nchez-P\u00e9rez, H.J., Pryg, K., Shinners, J., Murray, N., Mu\u00f1oz-Jim\u00e9nez, S.G., Bencomo-Alerm, A., G\u00f3mez-Bustamante, A., and Jonap\u00e1-G\u00f3mez, L. (2018). Magnetic Nanoparticle-Based Biosensing Assay Quantitatively Enhances Acid-Fast Bacilli Count in Paucibacillary Pulmonary Tuberculosis. Biosensors, 8.","DOI":"10.3390\/bios8040128"},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Bhusal, N., Shrestha, S., Pote, N., and Alocilja, E.C. (2019). Nanoparticle-Based Biosensing of Tuberculosis, an Affordable and Practical Alternative to Current Methods. Biosensors, 9.","DOI":"10.3390\/bios9010001"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"144","DOI":"10.1186\/1556-276X-7-144","article-title":"Magnetic nanoparticles: preparation, physical properties, and applications in biomedicine","volume":"7","author":"Akbarzadeh","year":"2012","journal-title":"Nanoscale Res. Lett."},{"key":"ref_36","unstructured":"(2019, October 14). Tuberculosis (TB). Available online: https:\/\/www.who.int\/news-room\/fact-sheets\/detail\/tuberculosis."},{"key":"ref_37","unstructured":"(2019, December 12). gli_mycobacteriology_lab_manual_quadri.indd. 154. Available online: https:\/\/www.scribd.com\/document\/305681530\/gli-mycobacteriology-lab-manual-web-pdf."},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Mohan, A., Padiadpu, J., Baloni, P., and Chandra, N. (2015). Complete Genome Sequences of a Mycobacterium smegmatis Laboratory Strain (MC2 155) and Isoniazid-Resistant (4XR1\/R2) Mutant Strains. Genome Announc., 3.","DOI":"10.1128\/genomeA.01520-14"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"68","DOI":"10.1016\/S0966-842X(01)02296-X","article-title":"Mycobacterium smegmatis and tuberculosis","volume":"10","author":"Tyagi","year":"2002","journal-title":"Trends Microbiol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"5133","DOI":"10.1074\/jbc.M610183200","article-title":"The Mycobacterium tuberculosis Cell-surface Glycoprotein Apa as a Potential Adhesin to Colonize Target Cells via the Innate Immune System Pulmonary C-type Lectin Surfactant Protein A","volume":"282","author":"Ragas","year":"2007","journal-title":"J. Biol. Chem."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1038\/ncomms5730","article-title":"Mycobacterium tuberculosis acquires iron by cell-surface sequestration and internalization of human holo-transferrin","volume":"5","author":"Boradia","year":"2014","journal-title":"Nat. Commun."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"5676","DOI":"10.1128\/IAI.72.10.5676-5686.2004","article-title":"The Glycan-Rich Outer Layer of the Cell Wall of Mycobacterium tuberculosis Acts as an Antiphagocytic Capsule Limiting the Association of the Bacterium with Macrophages","volume":"72","author":"Stokes","year":"2004","journal-title":"Infect. Immun."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"131","DOI":"10.1016\/S0065-2911(08)60016-8","article-title":"The Envelope Layers of Mycobacteria with Reference to their Pathogenicity","volume":"Volume 39","author":"Poole","year":"1997","journal-title":"Advances in Microbial Physiology"},{"key":"ref_44","unstructured":"(2019, October 25). Incidence of tuberculosis (per 100,000 people) | Data. Available online: https:\/\/data.worldbank.org\/indicator\/SH.TBS.INCD?name_desc=true."},{"key":"ref_45","unstructured":"(2019, October 15). Figure 2. The slow road to diagnosis of pulmonary tuberculosis (TB)... . Available online: https:\/\/www.researchgate.net\/figure\/The-slow-road-to-diagnosis-of-pulmonary-tuberculosis-TB-when-sputum-smear-microscopy-is_fig7_281642259."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"669","DOI":"10.1046\/j.1365-2672.2000.01020.x","article-title":"The physiology and pathogenicity of Mycobacterium tuberculosis grown under controlled conditions in a defined medium","volume":"88","author":"James","year":"2000","journal-title":"J. Appl. Microbiol."},{"key":"ref_47","unstructured":"(2019, October 17). Figure 4. Diagnostic needs at different health system levels. Available online: https:\/\/www.researchgate.net\/figure\/Diagnostic-needs-at-different-health-system-levels_fig8_281642259."}],"container-title":["Diagnostics"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2075-4418\/9\/4\/222\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T13:42:08Z","timestamp":1760190128000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2075-4418\/9\/4\/222"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2019,12,13]]},"references-count":47,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2019,12]]}},"alternative-id":["diagnostics9040222"],"URL":"https:\/\/doi.org\/10.3390\/diagnostics9040222","relation":{},"ISSN":["2075-4418"],"issn-type":[{"value":"2075-4418","type":"electronic"}],"subject":[],"published":{"date-parts":[[2019,12,13]]}}}