{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,14]],"date-time":"2025-10-14T01:19:18Z","timestamp":1760404758807,"version":"build-2065373602"},"reference-count":40,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2018,7,27]],"date-time":"2018-07-27T00:00:00Z","timestamp":1532649600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Sensors"],"abstract":"<jats:p>Adenosine deaminase (ADA), able to catalyze the irreversible deamination of adenosine into inosine, can be found in almost all tissues and plays an important role in several diseases. In this work, we developed a label-free fluorescence method for the detection of adenosine deaminase activity and inhibition. In the presence of ADA, ATP has been shown to be hydrolyzed. The ATP aptamer was shown to form a G-quadruplex\/thioflavin T (ThT) complex with ThT and exhibited an obvious fluorescence signal. However, the ATP aptamer could bind with ATP and exhibited a low fluorescence signal because of the absence of ADA. This assay showed high sensitivity to ADA with a detection limit of 1 U\/L based on an SNR of 3 and got a good linear relationship within the range of 1\u2013100 U\/L with R2 = 0.9909. The LOD is lower than ADA cutoff value (4 U\/L) in the clinical requirement and more sensitive than most of the reported methods. This technique exhibited high selectivity for ADA against hoGG I, UDG, RNase H and \u03bbexo. Moreover, this strategy was successfully applied for assaying the inhibition of ADA using erythro-9-(2-hydroxy-3-nonyl) adenine (EHNA) and, as such, demonstrated great potential for the future use in the diagnosis of ADA-relevant diseases, particularly in advanced drug development.<\/jats:p>","DOI":"10.3390\/s18082441","type":"journal-article","created":{"date-parts":[[2018,7,27]],"date-time":"2018-07-27T12:20:03Z","timestamp":1532694003000},"page":"2441","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["A Label-Free Fluorescent Assay for the Rapid and Sensitive Detection of Adenosine Deaminase Activity and Inhibition"],"prefix":"10.3390","volume":"18","author":[{"given":"Xinxing","family":"Tang","sequence":"first","affiliation":[{"name":"School of Art and Design, Changsha University of Science and Technology, Changsha 410114, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kefeng","family":"Wu","sequence":"additional","affiliation":[{"name":"School of Life Sciences, Central South University, Changsha 410013, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Han","family":"Zhao","sequence":"additional","affiliation":[{"name":"School of Life Sciences, Central South University, Changsha 410013, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mingjian","family":"Chen","sequence":"additional","affiliation":[{"name":"School of Life Sciences, Central South University, Changsha 410013, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Changbei","family":"Ma","sequence":"additional","affiliation":[{"name":"School of Life Sciences, Central South University, Changsha 410013, China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2018,7,27]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"5448","DOI":"10.1016\/S0021-9258(17)33080-6","article-title":"Human adenosine deaminase","volume":"251","author":"Kelley","year":"1976","journal-title":"J. Biol. Chem."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"479","DOI":"10.1042\/bj0330479","article-title":"The deaminase of adenosine and adenylic acid in blood and tissues","volume":"33","author":"Conway","year":"1939","journal-title":"Biochem. J."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"523","DOI":"10.1016\/j.neuint.2012.02.008","article-title":"Novel metabolic aspects related to adenosine deaminase inhibition in a human astrocytoma cell line","volume":"60","author":"Tozzi","year":"2012","journal-title":"Neurochem. Int."},{"key":"ref_4","first-page":"79","article-title":"Role of adenosine deaminase estimation in differentiation of tuberculous and non-tuberculous exudative pleural effusions","volume":"2","author":"Gupta","year":"2010","journal-title":"J. Clin. Med. Res."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"254","DOI":"10.1136\/jcp.34.3.254","article-title":"Enzyme and membrane markers in leukaemia: Recent developments","volume":"34","author":"Hofbrand","year":"1981","journal-title":"J. Clin. Pathol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"97","DOI":"10.1016\/S0385-8146(87)80027-5","article-title":"Adenosine deaminase and purine phosphorylase activities in lymphocytes and red blood cells of patients with carcinoma of the larynx","volume":"14","author":"Gierek","year":"1987","journal-title":"Auris Nasus Larynx"},{"key":"ref_7","first-page":"416","article-title":"Activities of adenosine deaminase and 5\u2032-nucleotidase in cancereous and non-cancereous human gastric tissues","volume":"110","author":"Gocmen","year":"2009","journal-title":"Bratisl. Lek. Listy"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"336","DOI":"10.1111\/j.1469-1809.2006.00338.x","article-title":"Carrier frequency of a nonsense mutation in the adenosine deaminase (ADA) gene implies a high incidence of ADA-deficient severe combined immunodeficiency (SCID) in Somalia and a single, common haplotype indicates common ancestry","volume":"71","author":"Sanchez","year":"2007","journal-title":"Ann. Hum. Genet."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"371","DOI":"10.1016\/S0167-5699(97)01047-5","article-title":"SCID: The role of adenosine deaminase deficienc","volume":"18","author":"Resta","year":"1997","journal-title":"Immunol. Today"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"105","DOI":"10.1002\/1098-1128(200103)21:2<105::AID-MED1002>3.0.CO;2-U","article-title":"Adenosine deaminase: Functional implications and different classes of inhibitors","volume":"21","author":"Cristalli","year":"2001","journal-title":"Med. Res. Rev."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"447","DOI":"10.1056\/NEJMoa0805817","article-title":"Gene Therapy for Immunodeficiency due to Adenosine Deaminase Deficiency","volume":"360","author":"Malacarne","year":"2009","journal-title":"Engl. J. Med."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1001","DOI":"10.1172\/JCI112866","article-title":"Elevated adenosine deaminase activity and hereditary hemolytic anemia. Evidence for abnormal translational control of protein synthesis","volume":"79","author":"Chottiner","year":"1987","journal-title":"J. Clin. Investig."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"61","DOI":"10.1016\/j.bios.2012.04.037","article-title":"Graphene oxide based fluorescent aptasensor for adenosine deaminase detection using adenosine as the substrate","volume":"37","author":"Xing","year":"2012","journal-title":"Biosens. Bioelectron."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"775","DOI":"10.1146\/annurev.pharmtox.41.1.775","article-title":"Molecular Approach to Adenosine Receptors: Receptor-Mediated Mechanisms of Tissue Protection","volume":"41","author":"Linden","year":"2001","journal-title":"Annu. Rev. Pharmacol. Toxicol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"146","DOI":"10.1016\/j.jchromb.2005.06.006","article-title":"Merits of HPLC-based method over spectrophotometric method for assessing the kinetics and inhibition of mammalian adenosine deaminase","volume":"822","author":"Paul","year":"2005","journal-title":"J. Chromatogr. B"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"313","DOI":"10.1016\/0022-1759(84)90406-X","article-title":"A colorimetric assay for serial determination of adenosine deaminase activity in small lymphocyte populations","volume":"73","author":"Vielh","year":"1984","journal-title":"J. Immunol. Methods"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"3207","DOI":"10.1021\/ac902771k","article-title":"Strategy to Fabricate an Electrochemical Aptasensor: Application to the Assay of Adenosine Deaminase Activity","volume":"82","author":"Zhang","year":"2010","journal-title":"Anal. Chem."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"422","DOI":"10.1016\/j.bios.2016.08.079","article-title":"The aptamer DNA-templated fluorescence silver nanoclusters: ATP detection and preliminary mechanism investigation","volume":"87","author":"Xu","year":"2017","journal-title":"Biosens. Bioelectron."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1572","DOI":"10.1039\/C4AN02070G","article-title":"A gold nanoparticle-based label free colorimetric aptasensor for adenosine deaminase detection and inhibition assay","volume":"140","author":"Cheng","year":"2015","journal-title":"Analyst"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"2438","DOI":"10.1039\/c3an36826b","article-title":"Fluorescence sensing of adenosine deaminase based on adenosine induced self-assembly of aptamer structures","volume":"138","author":"Feng","year":"2013","journal-title":"Analyst"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"850","DOI":"10.1038\/355850a0","article-title":"Selection in vitro of single-stranded DNA molecules that fold into specific ligand-binding structures","volume":"355","author":"Ellington","year":"1992","journal-title":"Nature"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"505","DOI":"10.1126\/science.2200121","article-title":"Systematic evolution of ligands by exponential enrichment: RNA ligands to bacteriophage T4 DNA polymerase","volume":"249","author":"Tuerk","year":"1990","journal-title":"Science"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"109","DOI":"10.1007\/s00604-017-2628-y","article-title":"Colorimetric determination of the activity of alkaline phosphatase based on the use of Cu(II)-modulated G-quadruplex-based DNAzymes","volume":"185","author":"Tang","year":"2018","journal-title":"Microchim. Acta"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"26","DOI":"10.1016\/S1367-5931(97)80105-6","article-title":"DNA aptamers and DNA enzymes","volume":"1","author":"Breaker","year":"1997","journal-title":"Curr. Opin. Chem. Biol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"2842","DOI":"10.1021\/cr300468w","article-title":"Aptamers from cell-based selection for bioanalytical applications","volume":"113","author":"Tan","year":"2013","journal-title":"Chem. Rev."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"29","DOI":"10.1016\/j.bios.2012.03.034","article-title":"Label-free detection of kanamycin based on the aptamer-functionalized conducting polymer\/gold nanocomposite","volume":"36","author":"Zhu","year":"2012","journal-title":"Biosens. Bioelectron."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"820","DOI":"10.1126\/science.287.5454.820","article-title":"Adaptive Recognition by Nucleic Acid Aptamers","volume":"287","author":"Hermann","year":"2000","journal-title":"Science"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1151","DOI":"10.1007\/s00604-017-2113-7","article-title":"Amperometric aptasensor for ochratoxin A based on the use of a gold electrode modified with aptamer, complementary DNA, SWCNTs and the redox marker methylene blue","volume":"184","author":"Abnous","year":"2017","journal-title":"Microchim. Acta"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"171","DOI":"10.1016\/j.aca.2013.06.029","article-title":"A colorimetric method for protein assay via exonuclease III-assisted signal attenuation strategy and specific DNA\u2013protein interaction","volume":"788","author":"Gao","year":"2013","journal-title":"Anal. Chim. Acta"},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Song, M.S., Sekhon, S.S., Shin, W.R., Kim, H.C., Min, J., Ahn, J.Y., and Kim, Y.H. (2017). Detecting and Discriminating Shigella sonnei Using an Aptamer-Based Fluorescent Biosensor Platform. Molecules, 22.","DOI":"10.3390\/molecules22050825"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"347","DOI":"10.1007\/s00604-018-2885-4","article-title":"Fluorometric aptasensor of ochratoxin A based on the use of graphene oxide and RNase H-aided amplification","volume":"7","author":"Ma","year":"2018","journal-title":"Microchim. Acta"},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Wu, K., Ma, C., Zhao, H., He, H., and Chen, H. (2018). Label-free G-quadruplex aptamer fluorescence assay for Ochratoxin A using a thioflavin T probe. Toxins, 10.","DOI":"10.3390\/toxins10050198"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1016\/j.mcp.2016.10.010","article-title":"Label-free monitoring of DNA polymerase activity based on a thrombin-binding aptamer G-quadruplex","volume":"32","author":"Wang","year":"2017","journal-title":"Mol. Cell. Probe"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"420","DOI":"10.1016\/j.bios.2013.05.051","article-title":"Stable label-free fluorescent sensing of biothiols based on ThT direct inducing conformation-specific G-quadruplex","volume":"49","author":"Tong","year":"2013","journal-title":"Biosens. Bioelectron."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"811","DOI":"10.1016\/j.bios.2016.07.083","article-title":"Label-free thioflavin T\/G-quadruplex-based real-time strand displacement amplification for biosensing applications","volume":"86","author":"Du","year":"2016","journal-title":"Biosens. Bioelectron."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"653","DOI":"10.1016\/j.talanta.2017.01.027","article-title":"Label-free fluorescent assay of T4 polynucleotide kinase phosphatase activity based on G-quadruplexe-thioflavin T complex","volume":"165","author":"Zhao","year":"2017","journal-title":"Talanta"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"164","DOI":"10.1016\/j.saa.2016.12.033","article-title":"A facile label-free fluorescence aptasensor for rapid detection of ATP based on G-quadruplex formation","volume":"175","author":"Liu","year":"2017","journal-title":"Spectrochim. Acta A"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"175","DOI":"10.1016\/j.jpba.2018.02.045","article-title":"Rapid, reliable, and sensitive detection of adenosine deaminase activity by UHPLC-Q-Orbitrap HRMS and its application to inhibitory activity evaluation of traditional Chinese medicines","volume":"153","author":"Qi","year":"2018","journal-title":"J. Pharm. Biomed. Anal."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1192","DOI":"10.1039\/C4AN01963F","article-title":"Label-free aptasensor for adenosine deaminase sensing based on fluorescence turn-on","volume":"140","author":"Zeng","year":"2015","journal-title":"Analyst"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"344","DOI":"10.1080\/15321819.2014.899255","article-title":"A New Method to Fabricate an Electrochemical Aptasensor to Assay Adenosine Deaminase Concentration using an Assistance DNA","volume":"35","author":"Huang","year":"2014","journal-title":"J. Immunoass. Immunochem."}],"container-title":["Sensors"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1424-8220\/18\/8\/2441\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T15:14:41Z","timestamp":1760195681000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1424-8220\/18\/8\/2441"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2018,7,27]]},"references-count":40,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2018,8]]}},"alternative-id":["s18082441"],"URL":"https:\/\/doi.org\/10.3390\/s18082441","relation":{},"ISSN":["1424-8220"],"issn-type":[{"type":"electronic","value":"1424-8220"}],"subject":[],"published":{"date-parts":[[2018,7,27]]}}}