{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,18]],"date-time":"2026-09-18T04:57:41Z","timestamp":1789707461542,"version":"4.0.1"},"reference-count":34,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2022,1,21]],"date-time":"2022-01-21T00:00:00Z","timestamp":1642723200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"National Institute of Health","award":["R01 DK067859"],"award-info":[{"award-number":["R01 DK067859"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJNS"],"abstract":"<jats:p>All newborn screening (NBS) for mucopolysaccharidosis-I and -II (MPS-I and MPS-II) is carried out via the measurement of \u03b1-iduronidase (IDUA) and iduronate-2-sulfatase (IDS) enzymatic activity, respectively, in dried blood spots (DBS). The majority of low enzyme results are due to pseudodeficiencies, and data from recent MPS-II population screenings and studies from the Mayo Clinic show that the false positive rate can be dramatically reduced by the inclusion of a second-tier analysis of glycosaminoglycans (GAGs) in DBS as part of NBS. In the present study, which focused on MPS-II, we obtained newborn DBS from 17 patients with severe MPS-II, 1 with attenuated MPS-II, and 6 patients with various IDS pseudodeficiencies. These samples were submitted to two different GAG mass spectrometry analyses in a comparative study: (1) internal disaccharide biomarkers and (2) endogenous biomarkers. For both of these methods, the biomarker levels in six patients with pseudodeficiencies were below the range measured in MPS-II patients. One patient with attenuated MPS-II was not distinguishable from severe disease patients, but all MPS-II patients were distinguishable from the reference range using both methods. The minimal differential factor (lowest GAG marker level in MPS-II samples divided by highest level in the reference range of 60 random newborns) was 3.01-fold for the internal disaccharide method. The endogenous biomarker method demonstrated an improved minimum differential of 5.41-fold. The minimum differential factors between MPS-II patients and patients with pseudodeficiencies for the internal disaccharide and endogenous biomarker methods were 3.77-fold and 2.06-fold, respectively. This study supports use of the second-tier GAG analysis of newborn DBS, especially the endogenous disaccharide method, as part of NBS to reduce the false positive rate.<\/jats:p>","DOI":"10.3390\/ijns8010009","type":"journal-article","created":{"date-parts":[[2022,1,23]],"date-time":"2022-01-23T20:32:52Z","timestamp":1642969972000},"page":"9","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":30,"title":["Evaluation of Two Methods for Quantification of Glycosaminoglycan Biomarkers in Newborn Dried Blood Spots from Patients with Severe and Attenuated Mucopolysaccharidosis Type II"],"prefix":"10.3390","volume":"8","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-0097-4222","authenticated-orcid":false,"given":"Zackary M.","family":"Herbst","sequence":"first","affiliation":[{"name":"Department of Chemistry, University of Washington, Seattle, WA 98195, USA"},{"name":"Department of Medicine, University of Washington, Seattle, WA 98195, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1978-0855","authenticated-orcid":false,"given":"Leslie","family":"Urdaneta","sequence":"additional","affiliation":[{"name":"National MPS Society, P.O. Box 14686, Durham, NC 27707-4686, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7154-4342","authenticated-orcid":false,"given":"Terri","family":"Klein","sequence":"additional","affiliation":[{"name":"National MPS Society, P.O. Box 14686, Durham, NC 27707-4686, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Barbara K.","family":"Burton","sequence":"additional","affiliation":[{"name":"Genetics, Birth Defects & Metabolism at Ann & Robert H. Lurie Children\u2019s Hospital of Chicago, Chicago, IL 60611, USA"},{"name":"Department of Pediatrics, Feinberg School of Medicine, Northwestern University, Chicago, IL 60611, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Khaja","family":"Basheeruddin","sequence":"additional","affiliation":[{"name":"Newborn Screening Laboratory, Illinois Department of Public Health, Chicago, IL 60604, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hsuan-Chieh","family":"Liao","sequence":"additional","affiliation":[{"name":"Department of Laboratory Medicine and Pathology, University of Washington, Seattle, WA 98195, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9092-8942","authenticated-orcid":false,"given":"Maria","family":"Fuller","sequence":"additional","affiliation":[{"name":"Genetics and Molecular Pathology, SA Pathology at Women\u2019s and Children\u2019s Hospital, North Adelaide 5006, Australia"},{"name":"School of Medicine, University of Adelaide, Adelaide 5005, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michael H.","family":"Gelb","sequence":"additional","affiliation":[{"name":"Department of Chemistry, University of Washington, Seattle, WA 98195, USA"},{"name":"Department of Biochemistry, University of Washington, Seattle, WA 98195, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,1,21]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"84","DOI":"10.1186\/s13023-018-0816-4","article-title":"Status of newborn screening and follow up investigations for Mucopolysaccharidoses I and II in Taiwan","volume":"13","author":"Chuang","year":"2018","journal-title":"Orphanet J. Rare Dis."},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Burton, B.K., Hickey, R., and Hitchins, L. (2020). Newborn Screening for Mucopolysaccharidosis Type II in Illinois: An Update. Int. J. Neonatal Screen., 6.","DOI":"10.3390\/ijns6030073"},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Bilyeu, H., Washburn, J., Vermette, L., and Klug, T. (2020). Validation and Implementation of a Highly Sensitive and Efficient Newborn Screening Assay for Mucopolysaccharidosis Type II. Int. J. Neonatal Screen., 6.","DOI":"10.3390\/ijns6040079"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"392","DOI":"10.1111\/j.1399-0004.1986.tb01896.x","article-title":"Full expression of Hunter\u2019s disease in a female with an X-chromosome deletion leading to non-random inactivation","volume":"30","author":"Broadhead","year":"1986","journal-title":"Clin. Genet."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"355","DOI":"10.1111\/j.1399-0004.1990.tb03519.x","article-title":"Hunter disease (mucopolysaccharidosis type II) in a karyotypically normal girl","volume":"37","author":"Clarke","year":"1990","journal-title":"Clin. Genet."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"361","DOI":"10.1002\/(SICI)1098-1004(1997)10:5<361::AID-HUMU5>3.0.CO;2-I","article-title":"Hunter disease in a girl caused by R468Q mutation in the iduronate-2-sulfatase gene and skewed inactivation of the X chromosome carrying the normal allele","volume":"10","author":"Sukegawa","year":"1997","journal-title":"Hum. Mutat."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"96","DOI":"10.1111\/j.1399-0004.1998.tb02654.x","article-title":"Brother\/sister siblings affected with Hunter disease: Evidence for skewed X chromosome inactivation","volume":"53","author":"Sukegawa","year":"1998","journal-title":"Clin. Genet."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"459","DOI":"10.1111\/j.1399-0004.2010.01574.x","article-title":"Female Hunter syndrome caused by a single mutation and familial XCI skewing: Implications for other X-linked disorders","volume":"80","author":"Kloska","year":"2011","journal-title":"Clin. Genet."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"450","DOI":"10.1002\/ajmg.a.34415","article-title":"Mucopolysaccharidosis type II in females and response to enzyme replacement therapy","volume":"158A","author":"Jurecka","year":"2012","journal-title":"Am. J. Med. Genet. A"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"2627","DOI":"10.1002\/ajmg.a.36667","article-title":"Mucopolysaccharidosis type II in a female patient with a reciprocal X;9 translocation and skewed X chromosome inactivation","volume":"164A","author":"Lonardo","year":"2014","journal-title":"Am. J. Med. Genet. A"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"557","DOI":"10.1038\/s10038-020-0744-8","article-title":"Newborn screening of mucopolysaccharidoses: Past, present, and future","volume":"65","author":"Arunkumar","year":"2020","journal-title":"J. Hum. Genet."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"204","DOI":"10.1016\/j.jpeds.2019.09.036","article-title":"Newborn screening for mucopolysaccharidoses: Results of a pilot study with 100,000 dried blood spots","volume":"216","author":"Scott","year":"2020","journal-title":"J. Pediatr."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"847","DOI":"10.1038\/gim.2017.194","article-title":"Precision newborn screening for lysosomal disorders","volume":"20","author":"Stoway","year":"2018","journal-title":"Genet. Med."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1108","DOI":"10.1038\/s41436-020-0764-y","article-title":"The critical role of psychosine in screening, diagnosis, and monitoring of Krabbe disease","volume":"22","author":"Guenzel","year":"2020","journal-title":"Genet. Med."},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Peck, D.S., Lacey, J.M., White, A.L., Pino, G., Studinski, A.L., Fisher, R., Ahmad, A., Spencer, L., Viall, S., and Shallow, N. (2020). Incorporation of Second-Tier Biomarker Testing Improves the Specificity of Newborn Screening for Mucopolysaccharidosis Type I. Int. J. Neonatal Screen., 6.","DOI":"10.3390\/ijns6010010"},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Herbst, Z.M., Urdaneta, L., Klein, T., Fuller, M., and Gelb, M.H. (2020). Evaluation of Multiple Methods for Quantification of Glycosaminoglycan Biomarkers in Newborn Dried Blood Spots from Patients with Severe and Attenuated Mucopolysaccharidosis-I. Int. J. Neonatal Screen., 6.","DOI":"10.3390\/ijns6030069"},{"key":"ref_17","first-page":"006","article-title":"Assay for Glycosaminoglycans by Tandem Mass Spectrometry and its Applications","volume":"2014","author":"Tomatsu","year":"2014","journal-title":"J. Anal. Bioanal. Tech."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"73","DOI":"10.1016\/j.ymgme.2013.07.016","article-title":"Glycan-based biomarkers for mucopolysaccharidoses","volume":"111","author":"Lawrence","year":"2014","journal-title":"Mol. Genet. Metab."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"753","DOI":"10.1038\/s41436-018-0136-z","article-title":"Disease and subtype specific signatures enable precise diagnosis of the mucopolysaccharidoses","volume":"21","author":"Saville","year":"2019","journal-title":"Genet. Med."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"291","DOI":"10.1038\/nmeth0408-291","article-title":"Disaccharide structure code for the easy representation of constituent oligosaccharides from glycosaminoglycans","volume":"5","author":"Lawrence","year":"2008","journal-title":"Nat. Methods"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"33674","DOI":"10.1074\/jbc.M804288200","article-title":"Evolutionary differences in glycosaminoglycan fine structure detected by quantitative glycan reductive isotope labeling","volume":"283","author":"Lawrence","year":"2008","journal-title":"J. Biol. Chem."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"733","DOI":"10.1203\/01.PDR.0000141987.69757.DD","article-title":"Disease-specific markers for the mucopolysaccharidoses","volume":"56","author":"Fuller","year":"2004","journal-title":"Pediatr. Res."},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Herbst, Z.M., Turgeon, C.M., Biski, C., Khaledi, H., Shoemaker, N.B., DeArmond, P.D., Smith, S., Orsini, J., Matern, D., and Gelb, M.H. (2020). Achieving Congruence among Reference Laboratories for Absolute Abundance Measurement of Analytes for Rare Diseases: Psychosine for Diagnosis and Prognosis of Krabbe Disease. Int. J. Neonatal Screen., 6.","DOI":"10.3390\/ijns6020029"},{"key":"ref_24","first-page":"100563","article-title":"Newborn screening for mucopolysaccharidoses: Measurement of glycosaminoglycans by LC-MS\/MS","volume":"22","author":"Stapleton","year":"2020","journal-title":"Mol. Genet. Metab. Rep."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"705","DOI":"10.1016\/j.ymgme.2012.09.024","article-title":"Heparan sulfate and dermatan sulfate derived disaccharides are sensitive markers for newborn screening for mucopolysaccharidoses types I, II and III","volume":"107","author":"Wagemans","year":"2012","journal-title":"Mol. Genet. Metab."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"247","DOI":"10.1016\/j.ymgme.2016.12.010","article-title":"Glycosaminoglycan levels in dried blood spots of patients with mucopolysaccharidoses and mucolipidoses","volume":"120","author":"Kubaski","year":"2017","journal-title":"Mol. Genet. Metab."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1779","DOI":"10.1007\/s00216-021-03152-7","article-title":"Stability and recovery issues concerning chondroitin sulfate disaccharide analysis","volume":"413","author":"Drahos","year":"2021","journal-title":"Anal. Bioanal. Chem."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"771","DOI":"10.1016\/j.cca.2012.01.012","article-title":"An improved method for glycosaminoglycan analysis by LC-MS\/MS of urine samples collected on filter paper","volume":"413","author":"Lavoie","year":"2012","journal-title":"Clin. Chim. Acta"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"151","DOI":"10.1007\/s10545-016-9981-6","article-title":"Newborn screening for mucopolysaccharidoses: A pilot study of measurement of glycosaminoglycans by tandem mass spectrometry","volume":"40","author":"Kubaski","year":"2017","journal-title":"J. Inherit. Metab. Dis."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"560442","DOI":"10.3389\/fcell.2020.560442","article-title":"Research and Application of Chondroitin Sulfate\/Dermatan Sulfate-Degrading Enzymes","volume":"8","author":"Wang","year":"2020","journal-title":"Front. Cell Dev. Biol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"330","DOI":"10.1016\/0003-2697(62)90095-7","article-title":"A modified uronic acid carbazole reaction","volume":"4","author":"Bitter","year":"1962","journal-title":"Anal. Biochem."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"343","DOI":"10.1042\/bj1310343","article-title":"The quantitative measurement of Alcian Blue-glycosaminoglycan complexes","volume":"131","author":"Whiteman","year":"1973","journal-title":"Biochem. J."},{"key":"ref_33","first-page":"419","article-title":"Clinical application of a new simple method for the identification of mucopolysaccharidoses","volume":"42","author":"Coppa","year":"1987","journal-title":"Helv. Paediatr. Acta"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"147","DOI":"10.1016\/0009-8981(85)90318-3","article-title":"Screening test for urinary glycosaminoglycans and differentiation of various mucopolysaccharidoses","volume":"151","author":"Huang","year":"1985","journal-title":"Clin. Chim. Acta"}],"container-title":["International Journal of Neonatal Screening"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2409-515X\/8\/1\/9\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T22:05:44Z","timestamp":1760133944000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2409-515X\/8\/1\/9"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,1,21]]},"references-count":34,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2022,3]]}},"alternative-id":["ijns8010009"],"URL":"https:\/\/doi.org\/10.3390\/ijns8010009","relation":{},"ISSN":["2409-515X"],"issn-type":[{"value":"2409-515X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,1,21]]}}}