{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,4]],"date-time":"2026-08-04T00:34:52Z","timestamp":1785803692139,"version":"3.56.0"},"reference-count":13,"publisher":"MDPI AG","issue":"18","license":[{"start":{"date-parts":[[2021,9,21]],"date-time":"2021-09-21T00:00:00Z","timestamp":1632182400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Italian Ministry of Health","award":["RF-2019-12370600"],"award-info":[{"award-number":["RF-2019-12370600"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecules"],"abstract":"<jats:p>Adenosine Deaminase 2 Deficiency (DADA2) (OMIM: 607575) is a monogenic, autoinflammatory disease caused by the loss of functional homozygous or heterozygous mutations in the ADA 2 gene (previously CECR1, Cat Eye Syndrome Chromosome Region 1). A timely diagnosis is crucial to start Anti-TNF therapies that are efficacious in controlling the disease. The confirmation of DADA2 is based on DNA sequencing and enzymatic assay. It is, thus, very important to have robust and reliable assays that can be rapidly utilized in specialized laboratories that can centralize samples from other centers. In this paper, we show a novel enzymatic assay based on liquid chromatography-tandem mass spectrometry that allows the accurate determination of the ADA2 enzyme activity starting from very small amounts of plasma spotted on filter paper (dried plasma spot). The method allows significantly distinguishing healthy controls from affected patients and carriers and could be of help in implementing the diagnostic workflow of DADA2.<\/jats:p>","DOI":"10.3390\/molecules26185707","type":"journal-article","created":{"date-parts":[[2021,9,21]],"date-time":"2021-09-21T22:34:01Z","timestamp":1632263641000},"page":"5707","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":16,"title":["A Novel LC\u2013MS\/MS-Based Method for the Diagnosis of ADA2 Deficiency from Dried Plasma Spot"],"prefix":"10.3390","volume":"26","author":[{"given":"Alessia","family":"Cafaro","sequence":"first","affiliation":[{"name":"Chromatography and Mass Spectrometry Section, Central Laboratory of Analysis, IRCCS Istituto Giannina Gaslini, 16147 Genoa, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Federica","family":"Pigliasco","sequence":"additional","affiliation":[{"name":"Chromatography and Mass Spectrometry Section, Central Laboratory of Analysis, IRCCS Istituto Giannina Gaslini, 16147 Genoa, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1148-427X","authenticated-orcid":false,"given":"Sebastiano","family":"Barco","sequence":"additional","affiliation":[{"name":"Chromatography and Mass Spectrometry Section, Central Laboratory of Analysis, IRCCS Istituto Giannina Gaslini, 16147 Genoa, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Federica","family":"Penco","sequence":"additional","affiliation":[{"name":"Center for Autoinflammatory Diseases and Immunodeficiencies, IRCCS Istituto Giannina Gaslini, 16147 Genoa, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5868-7699","authenticated-orcid":false,"given":"Francesca","family":"Schena","sequence":"additional","affiliation":[{"name":"Center for Autoinflammatory Diseases and Immunodeficiencies, IRCCS Istituto Giannina Gaslini, 16147 Genoa, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0131-7846","authenticated-orcid":false,"given":"Roberta","family":"Caorsi","sequence":"additional","affiliation":[{"name":"Center for Autoinflammatory Diseases and Immunodeficiencies, IRCCS Istituto Giannina Gaslini, 16147 Genoa, Italy"},{"name":"Clinics of Pediatrics and Rheumatology, IRCCS Istituto Giannina Gaslini, 16147 Genoa, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7129-868X","authenticated-orcid":false,"given":"Stefano","family":"Volpi","sequence":"additional","affiliation":[{"name":"Center for Autoinflammatory Diseases and Immunodeficiencies, IRCCS Istituto Giannina Gaslini, 16147 Genoa, Italy"},{"name":"Dipartimento di Neuroscienze, Riabilitazione, Oftalmologia, Genetica e Scienze Materno-Infantili, University of Genoa, 16132 Genoa, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gino","family":"Tripodi","sequence":"additional","affiliation":[{"name":"Chromatography and Mass Spectrometry Section, Central Laboratory of Analysis, IRCCS Istituto Giannina Gaslini, 16147 Genoa, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marco","family":"Gattorno","sequence":"additional","affiliation":[{"name":"Center for Autoinflammatory Diseases and Immunodeficiencies, IRCCS Istituto Giannina Gaslini, 16147 Genoa, Italy"},{"name":"Clinics of Pediatrics and Rheumatology, IRCCS Istituto Giannina Gaslini, 16147 Genoa, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Giuliana","family":"Cangemi","sequence":"additional","affiliation":[{"name":"Chromatography and Mass Spectrometry Section, Central Laboratory of Analysis, IRCCS Istituto Giannina Gaslini, 16147 Genoa, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,9,21]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1186\/s12969-016-0111-7","article-title":"Monogenic polyarteritis: The lesson of ADA2 deficiency","volume":"14","author":"Caorsi","year":"2016","journal-title":"Pediatr. Rheumatol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1664","DOI":"10.1016\/j.jaci.2019.12.908","article-title":"Genotype and functional correlates of disease phenotype in deficiency of adenosine deaminase 2 (DADA2)","volume":"145","author":"Lee","year":"2020","journal-title":"J. Allergy Clin. Immunol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"206","DOI":"10.1002\/eji.202048549","article-title":"Dysregulation in B-cell responses and T follicular helper cell function in ADA2 deficiency patients","volume":"51","author":"Schena","year":"2021","journal-title":"Eur. J. Immunol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1648","DOI":"10.1136\/annrheumdis-2016-210802","article-title":"ADA2 deficiency (DADA2) as an unrecognised cause of early onset polyarteritis nodosa and stroke: A multicentre national study","volume":"76","author":"Caorsi","year":"2017","journal-title":"Ann. Rheum. Dis."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"51","DOI":"10.1042\/BJ20050683","article-title":"Human ADA2 belongs to a new family of growth factors with adenosine deaminase activity","volume":"391","author":"Zavialov","year":"2005","journal-title":"Biochem. J."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"911","DOI":"10.1056\/NEJMoa1307361","article-title":"Early-Onset Stroke and Vasculopathy Associated with Mutations in ADA2","volume":"370","author":"Zhou","year":"2014","journal-title":"N. Engl. J. Med."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"921","DOI":"10.1056\/NEJMoa1307362","article-title":"Mutant Adenosine Deaminase 2 in a Polyarteritis Nodosa Vasculopathy","volume":"370","author":"Pierce","year":"2014","journal-title":"N. Engl. J. Med."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"512","DOI":"10.1002\/art.41549","article-title":"Sequence-Based Screening of Patients With Idiopathic Polyarteritis Nodosa, Granulomatosis With Polyangiitis, and Microscopic Polyangiitis for Deleterious Genetic Variants in ADA2","volume":"73","author":"Schnappauf","year":"2021","journal-title":"Arthritis Rheumatol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"316","DOI":"10.1016\/j.jpeds.2016.06.058","article-title":"Extending the Clinical Phenotype of Adenosine Deaminase 2 Deficiency","volume":"177","author":"Brooks","year":"2016","journal-title":"J. Pediatr."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"114292","DOI":"10.1016\/j.ab.2021.114292","article-title":"Enzyme activity in dried blood spot as a diagnostic tool for adenosine deaminase 2 deficiency","volume":"628","author":"Ito","year":"2021","journal-title":"Anal. Biochem."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1582","DOI":"10.1056\/NEJMc1801927","article-title":"Treatment Strategies for Deficiency of Adenosine Deaminase 2","volume":"380","author":"Ombrello","year":"2019","journal-title":"New Engl. J. of. Med."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"732","DOI":"10.1111\/cge.13518","article-title":"ADA2 deficiency due to a novel structural variation in 22q11.1","volume":"95","author":"Grossi","year":"2019","journal-title":"Clin. Genet."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"41","DOI":"10.1016\/j.pisc.2014.02.005","article-title":"Enzyme assays","volume":"1","author":"Bisswanger","year":"2014","journal-title":"Perspect. Sci."}],"container-title":["Molecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1420-3049\/26\/18\/5707\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T07:02:44Z","timestamp":1760166164000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1420-3049\/26\/18\/5707"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,9,21]]},"references-count":13,"journal-issue":{"issue":"18","published-online":{"date-parts":[[2021,9]]}},"alternative-id":["molecules26185707"],"URL":"https:\/\/doi.org\/10.3390\/molecules26185707","relation":{},"ISSN":["1420-3049"],"issn-type":[{"value":"1420-3049","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,9,21]]}}}