title: Brazilian reference values for MPS II screening in dried blood spots--a fluorimetric assay.
abstract: OBJECTIVE
  Mucopolysaccharidosis II (MPS II), or Hunter Syndrome, is a lysosomal storage disorder that is caused by the deficiency or absence of iduronate-2-sulfatase (IDS) enzyme; in this disease, early diagnosis is essential to provide higher life expectancy for patients. This study validates a fluorimetric assay that is used to assess IDS enzyme activity using dried blood spot (DBS) samples and presents the reference interval for the Brazilian population.
  
  
  METHODS
  Venous blood sample was collected in heparin tubes for leukocyte extraction and DBS preparation. IDS activity in the leukocytes was analyzed, and the results were considered the gold standard reference for the categorization of volunteers as positive or negative controls (PC and NC, respectively). IDS activity in the DBS was analyzed using an adapted version of the leukocyte assay. Statistical analyses were performed using a ROC curve to determine cutoff values and using a parametric Student's t test to compare values between genders. To verify that the assay yielded consistent results, a Bland-Altman plot was prepared.
  
  
  RESULTS
  Leukocyte IDS activity values ranged between 2.71 and 17.36 nmol/mg protein/h in the NC group and between 0 and 0.11 nmol/mg protein/h in the PC group. Based on the DBS assay, activities ranged between 1.83 and 16.86 μmol/L blood/h in the NC group and between 0.58 and 4.32 μmol/L blood/h in the PC group.
  
  
  CONCLUSIONS
  Reference values of IDS activity were determined in DBS with acceptable sensitivity and specificity. Therefore, the DBS assay described in this work may be a useful tool to screen MPS II patients in the Brazilian population.
authors: Laboratório de Erros Inatos do Metabolismo, Department of Psychobiology, Universidade Federal de São Paulo, Rua Napoleão de Barros, 925, 3rd Floor, 04024-002 São Paulo, SP, Brazil. Electronic address: marina.mrez@gmail.com.
authors: Marina Mastelaro
authors: MM
authors: Rezende
authors: Department of Pediatrics, Universidade Federal de São Paulo, Rua Botucatu, 598, 04023-062 São Paulo, SP, Brazil. Electronic address: kbmuller@gmail.com.
authors: Karen Barbosa
authors: KB
authors: Müller
authors: Laboratório de Erros Inatos do Metabolismo, Department of Psychobiology, Universidade Federal de São Paulo, Rua Napoleão de Barros, 925, 3rd Floor, 04024-002 São Paulo, SP, Brazil. Electronic address: nessagp@yahoo.com.br.
authors: Vanessa Gonçalves
authors: VG
authors: Pereira
authors: Laboratório de Erros Inatos do Metabolismo, Department of Psychobiology, Universidade Federal de São Paulo, Rua Napoleão de Barros, 925, 3rd Floor, 04024-002 São Paulo, SP, Brazil. Electronic address: vaniadalmeida@uol.com.br.
authors: Vânia
authors: V
authors: D'Almeida
mesh_terms: D000293:Adolescent; D001681:Biological Assay / Q000379:methods; D001938:Brazil; D059788:Dried Blood Spot Testing / Q000379:methods; D005260:Female; D005470:Fluorometry / Q000379:methods; D006801:Humans; D007962:Leukocytes / Q000378:metabolism; D008297:Male; D016532:Mucopolysaccharidosis II / Q000097:blood* / Q000175:diagnosis* / Q000378:metabolism; D012016:Reference Values; D012680:Sensitivity and Specificity
keywords: Dried blood spot; Fluorimetric assay; Hunter Syndrome; Leukocyte; Mucopolysaccharidosis II