{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,11,4]],"date-time":"2026-11-04T18:08:56Z","timestamp":1793815736984,"version":"4.6.1"},"reference-count":29,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2025,3,3]],"date-time":"2025-03-03T00:00:00Z","timestamp":1740960000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Hunan Provincial Natural Science Foundation of China","award":["2022JJ30229"],"award-info":[{"award-number":["2022JJ30229"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biosensors"],"abstract":"<jats:p>The accurate detection of Mycobacterium tuberculosis (MTB) is a pressing challenge in the precise prevention and control of tuberculosis. Currently, the efficiency and accuracy of drug resistance detection for MTB are low, and cross-contamination is common, making it inadequate for clinical needs. This study developed a rapid nucleic acid detection method for MTB based on scattering loop-mediated isothermal amplification (LAMP). Specific primers for the MTB-specific gene (Ag85B) were designed, and the LAMP reaction system was optimized using a self-developed scattering LAMP turbidimeter. Experimental results showed that the optimal reaction system included 1.5 \u00b5L of 100 mmol\/L magnesium ions, 3.5 \u00b5L of 10 mmol\/L dNTPs, 6 \u00b5L of 1.6 mol\/L betaine, and a reaction temperature of 65 \u00b0C. The minimum detection limit was 12.40 ng\/L, with the fastest detection time being approximately 10 min. The reaction exhibited good specificity, with no amplification bands for other pathogens. Twenty culture-positive samples and twenty culture-negative samples were tested in parallel; the accuracy of the positive group was 100%, the detection time was (24.9 \u00b1 13 min), and there was no negative detection. This method features high detection efficiency, low cost, high accuracy, and effectively reduces cross-contamination, providing a new technology for the rapid clinical detection of MTB.<\/jats:p>","DOI":"10.3390\/bios15030162","type":"journal-article","created":{"date-parts":[[2025,3,4]],"date-time":"2025-03-04T03:26:21Z","timestamp":1741058781000},"page":"162","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["A Novel Rapid Detection Method for Mycobacterium tuberculosis Based on Scattering-Light Turbidity Using Loop-Mediated Isothermal Amplification"],"prefix":"10.3390","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0009-0001-8150-9284","authenticated-orcid":false,"given":"Meimei","family":"Zeng","sequence":"first","affiliation":[{"name":"Hunan Key Laboratory of Biomedical Nanomaterials and Devices, Hunan University of Technology, Zhuzhou 412007, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xinru","family":"Wang","sequence":"additional","affiliation":[{"name":"Hunan Key Laboratory of Biomedical Nanomaterials and Devices, Hunan University of Technology, Zhuzhou 412007, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zifeng","family":"Tan","sequence":"additional","affiliation":[{"name":"Hunan Key Laboratory of Biomedical Nanomaterials and Devices, Hunan University of Technology, Zhuzhou 412007, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wenyan","family":"Guo","sequence":"additional","affiliation":[{"name":"Hunan Key Laboratory of Biomedical Nanomaterials and Devices, Hunan University of Technology, Zhuzhou 412007, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yan","family":"Deng","sequence":"additional","affiliation":[{"name":"MOE Key Lab of Rare Pediatric Diseases, Hengyang Medical College, University of South China, Hengyang 421001, China"},{"name":"Institute for Future Sciences, University of South China, Changsha 410008, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Song","family":"Li","sequence":"additional","affiliation":[{"name":"MOE Key Lab of Rare Pediatric Diseases, Hengyang Medical College, University of South China, Hengyang 421001, China"},{"name":"Institute for Future Sciences, University of South China, Changsha 410008, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Libo","family":"Nie","sequence":"additional","affiliation":[{"name":"Hunan Key Laboratory of Biomedical Nanomaterials and Devices, Hunan University of Technology, Zhuzhou 412007, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nongyue","family":"He","sequence":"additional","affiliation":[{"name":"Hunan Key Laboratory of Biomedical Nanomaterials and Devices, Hunan University of Technology, Zhuzhou 412007, China"},{"name":"State Key Laboratory of Digital Medical Engineering, School of Biological Science and Medical Engineering, Southeast University, Nanjing 210096, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1246-3390","authenticated-orcid":false,"given":"Zhu","family":"Chen","sequence":"additional","affiliation":[{"name":"Hunan Key Laboratory of Biomedical Nanomaterials and Devices, Hunan University of Technology, Zhuzhou 412007, China"},{"name":"MOE Key Lab of Rare Pediatric Diseases, Hengyang Medical College, University of South China, Hengyang 421001, China"},{"name":"Institute for Future Sciences, University of South China, Changsha 410008, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,3,3]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"e25122","DOI":"10.1002\/jcla.25122","article-title":"Mycobacterium tuberculosis: The Mechanism of Pathogenicity, Immune Responses, and Diagnostic Challenges","volume":"38","author":"Mohammadnabi","year":"2024","journal-title":"J. Clin. Lab. Anal."},{"key":"ref_2","unstructured":"World Health Organization (2024). Global Tuberculosis Report 2024, World Health Organization."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"617","DOI":"10.1038\/s41579-024-01025-1","article-title":"Drug-resistant tuberculosis: A persistent global health concern","volume":"22","author":"Farhat","year":"2024","journal-title":"Nat. Rev. Microbiol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"e9","DOI":"10.1016\/j.cmi.2021.09.014","article-title":"Whole-genome sequencing for surveillance of tuberculosis drug resistance and determination of resistance level in China","volume":"28","author":"Liu","year":"2022","journal-title":"Clin. Microbiol. Infect."},{"key":"ref_5","first-page":"000815","article-title":"Transmission, distribution and drug resistance-conferring mutations of extensively drug-resistant tuberculosis in the Western Cape Province, South Africa","volume":"8","author":"Oostvogels","year":"2022","journal-title":"Microb. Genom."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Wang, Y., Wang, C., and Zhou, Z. (2023). Advances in Simple, Rapid, and Contamination-Free Instantaneous Nucleic Acid Devices for Pathogen Detection. Biosensors, 13.","DOI":"10.3390\/bios13070732"},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Liebenberg, D., Gordhan, B.G., and Kana, B.D. (2022). Drug resistant tuberculosis: Implications for transmission, diagnosis, and disease management. Front. Cell. Infect. Microbiol., 12.","DOI":"10.3389\/fcimb.2022.943545"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"108504","DOI":"10.1016\/j.cclet.2023.108504","article-title":"High-throughput fluorescent screening of thioredoxin reductase inhibitors to inhibit Mycobacterium tuberculosis","volume":"35","author":"Yan","year":"2024","journal-title":"Chin. Chem. Lett."},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Rabaan, A.A., Mutair, A.A., and Albayat, H. (2022). Tools to alleviate the drug resistance in Mycobacterium tuberculosis. Molecules, 27.","DOI":"10.3390\/molecules27206985"},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Bernardini, R., Tengattini, S., and Li, Z. (2024). Effect of glycosylation on the affinity of the MTB protein Ag85B for specific antibodies: Towards the design of a dual-acting vaccine against tuberculosis. Biol. Direct, 19.","DOI":"10.1186\/s13062-024-00454-5"},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Dewi, D.N.S.S., Mertaniasih, N.M. (2023). Antibodies against native proteins of Mycobacterium tuberculosis can detect pulmonary tuberculosis patients. Sci. Rep., 13.","DOI":"10.1038\/s41598-023-39436-4"},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Garc\u00eda-Ruiz, V., Ordu\u00f1a, P., and Castillo-Rodal, A.I. (2022). Proteome Profile Changes Induced by Heterologous Overexpression of Mycobacterium tuberculosis-Derived Antigens PstS-1 (Rv0934) and Ag85B (Rv1886c) in Mycobacterium microti. Biomolecules, 12.","DOI":"10.3390\/biom12121836"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"62","DOI":"10.1093\/ajcp\/aqad012","article-title":"Detection of antigen Ag85B expression is useful for the diagnosis of tuberculosis, especially for those with an antituberculosis treatment history","volume":"160","author":"Dong","year":"2023","journal-title":"Am. J. Clin. Pathol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1782","DOI":"10.1007\/s00247-023-05650-5","article-title":"Imaging recommendations and algorithms for pediatric tuberculosis: Part 2-extrathoracic tuberculosis","volume":"53","author":"Laya","year":"2023","journal-title":"Pediatr. Radiol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"4682","DOI":"10.1016\/j.cell.2022.10.025","article-title":"Immune cell interactions in tuberculosis","volume":"185","author":"Flynn","year":"2022","journal-title":"Cell"},{"key":"ref_16","unstructured":"Wang, S., Zhang, Y., He, Y., Wu, D., and Tang, S. (2022). Development of a LAMP assay for rapid detection of Mycobacterium tuberculosis complex in clinical specimens. J. Microbiol. Methods, 192."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"e13698","DOI":"10.1111\/cpr.13698","article-title":"Single-cell sequencing: Current applications in various tuberculosis specimen types","volume":"57","author":"Zeng","year":"2024","journal-title":"Cell Prolif."},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Zouaghi, N., Aziz, S., Shah, I., Aamouche, A., Jung, D.W., Lakssir, B., and Ressami, E.M. (2023). Miniaturized rapid electrochemical immunosensor based on screen printed carbon electrodes for mycobacterium tuberculosis detection. Biosensors, 13.","DOI":"10.3390\/bios13060589"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"E63","DOI":"10.1093\/nar\/28.12.e63","article-title":"Loop-mediated isothermal amplification of DNA","volume":"28","author":"Notomi","year":"2000","journal-title":"Nucleic Acids Res."},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Park, J.W. (2022). Principles and Applications of Loop-Mediated Isothermal Amplification to Point-of-Care Tests. Biosensors, 10.","DOI":"10.3390\/bios12100857"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1933","DOI":"10.1016\/j.cclet.2021.11.051","article-title":"Highly sensitive smartphone-based detection of Listeria monocytogenes using SYTO9","volume":"33","author":"Liu","year":"2022","journal-title":"Chin. Chem. Lett."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"107701","DOI":"10.1016\/j.cclet.2022.07.044","article-title":"Novel aerosol detection platform for SARS-CoV-2: Based on specific magnetic nanoparticles adsorption sampling and digital droplet PCR detection","volume":"34","author":"Chen","year":"2023","journal-title":"Chin. Chem. Lett."},{"key":"ref_23","first-page":"122751","article-title":"A novel turbidimetric assay combined with loop-mediated isothermal amplification for sensitive nucleic acid detection","volume":"235","author":"Wu","year":"2021","journal-title":"Talanta"},{"key":"ref_24","unstructured":"Wei, W., Zhang, Y., Li, C., Smith, J., Johnson, K., Brown, L., Chen, H., Liu, Q., Yang, F., and Thompson, R. (2023). A microfluidic turbidimetric sensor for rapid nucleic acid detection. Biosens. Bioelectron., 208."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"123","DOI":"10.1007\/978-1-0716-3605-3_12","article-title":"Measuring of Alpha-1 Antitrypsin Concentration by Nephelometry or Turbidimetry","volume":"2750","year":"2024","journal-title":"Methods Mol. Biol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"159","DOI":"10.1016\/j.cclet.2019.05.003","article-title":"Asymmetric polymerase chain reaction and loop-mediated isothermal amplification (AP-LAMP) for ultrasensitive detection of microRNAs","volume":"31","author":"Wang","year":"2020","journal-title":"Chin. Chem. Lett."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"109308","DOI":"10.1016\/j.cclet.2023.109308","article-title":"Preparation of norovirus GII loop mediated isothermal amplification freeze-drying microsphere reagents and its application in an on-site integrated rapid detection platform","volume":"35","author":"Wu","year":"2024","journal-title":"Chin. Chem. Lett."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"6994","DOI":"10.1038\/s41467-021-27076-z","article-title":"Paper microfluidic implementation of loop mediated isothermal amplification for early diagnosis of hepatitis C virus","volume":"12","author":"Witkowska","year":"2021","journal-title":"Nat. Commun."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"4053","DOI":"10.1016\/j.cclet.2022.01.072","article-title":"Development and evaluation of a thermostatic nucleic acid testing device based on magnesium pyrophosphate precipitation for detecting Enterocytozoon hepatopenaei","volume":"33","author":"Chen","year":"2022","journal-title":"Chin. Chem. Lett."}],"container-title":["Biosensors"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2079-6374\/15\/3\/162\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T16:46:33Z","timestamp":1760028393000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2079-6374\/15\/3\/162"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,3,3]]},"references-count":29,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2025,3]]}},"alternative-id":["bios15030162"],"URL":"https:\/\/doi.org\/10.3390\/bios15030162","relation":{},"ISSN":["2079-6374"],"issn-type":[{"value":"2079-6374","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,3,3]]}}}