title: Effectiveness of calretinin and role of age in the diagnosis of Hirschsprung disease.
abstract: OBJECTIVE
  Gold standard in Hirschsprung disease (HD) diagnosis is rectal suction biopsy (RSB) with hematoxylin and eosin and acetylcholinesterase (ACE) stainings. Sensitivity is 83-93 % and specificity is between 95 and 99 %. The rate of non-conclusive results (NC) due to inadequate samples or variability in the interpretation is about 11-37.8 %. Interpretation is still difficult in neonates. Calretinin (C) is a calcium-binding protein, expressed in the nervous system. It has been introduced as a marker to improve the diagnosis accuracy in HD. We compare sensitivity, specificity between ACE and ACE + C and investigate whether the introduction of C could reduce the rate of inconclusive results. We also tried to define the most adequate age to obtain accurate results from RSB.
  
  
  METHODS
  Data from patients undergoing rectal suction biopsies from 2005 to 2014 was analyzed. Until 2010 we used ACE; since 2011 we used ACE + C. The ROC curve analysis of the NC results regarding the age, showed an optimal cut-off point at 1.5 months (m). It divides the sample into two groups which we compared.
  
  
  RESULTS
  We analyzed 91 patients. Results of the RSB: 40 HD (44 %), 34 no Hirschsprung (37.4 %) and 17 NC (18.7 %). Sensitivity = 97.5 %, specificity = 97.1 %, not including the NC (1 false positive, 1 false negative). Results depending on the staining: ACE (n = 58) (%) ACE + C (n = 33) (%) p Sensitivity 96 100 1 Specificity 94.7 100 1 NC 24.1 9.1 0.077 Results depending on the age: <1.5 m (n = 27) (%) >1.5 m (n = 64) (%) p Sensitivity 92.3 100 0.325 Specificity 100 96.8 1 NC 40.7 9.4 0.001 CONCLUSION: Calretinin decreases the rate of inconclusive results, but not significantly. The percentage of inconclusive results decreases in patients olders than 1.5 m. Further studies are necessary to determine if this technique is useful to improve RSB results in infants younger than 1.5 months.
authors.affiliation: Pediatric Surgery Department, Hospital Sant Joan de Déu, Universitat de Barcelona, Passeig Sant Joan de Déu 2, Esplugues de Llobregat, 08950, Barcelona, Spain.
authors.forename: Irene
authors.initials: I
authors.lastname: de Haro Jorge
authors.affiliation: Pediatric Surgery Department, Hospital Sant Joan de Déu, Universitat de Barcelona, Passeig Sant Joan de Déu 2, Esplugues de Llobregat, 08950, Barcelona, Spain.
authors.forename: Pedro
authors.initials: P
authors.lastname: Palazón Bellver
authors.affiliation: Pediatric Surgery Department, Hospital Sant Joan de Déu, Universitat de Barcelona, Passeig Sant Joan de Déu 2, Esplugues de Llobregat, 08950, Barcelona, Spain. vjulia@hsjdbcn.org.
authors.forename: Victoria
authors.initials: V
authors.lastname: Julia Masip
authors.affiliation: Pediatric Surgery Department, Hospital Sant Joan de Déu, Universitat de Barcelona, Passeig Sant Joan de Déu 2, Esplugues de Llobregat, 08950, Barcelona, Spain.
authors.forename: Laura
authors.initials: L
authors.lastname: Saura García
authors.affiliation: Pathology Department, Hospital Sant Joan de Déu, Universitat de Barcelona, Passeig Sant Joan de Déu 2, Esplugues de Llobregat, 08950, Barcelona, Spain.
authors.forename: Teresa
authors.initials: T
authors.lastname: Ribalta Farres
authors.affiliation: Parc Sanitari Sant Joan de Déu, Universitat de Barcelona, Carrer Antoni Pujadas 42, Sant Boi de Llobregat, 08830, Barcelona, Spain.
authors.forename: Daniel
authors.initials: D
authors.lastname: Cuadras Pallejà
authors.affiliation: Pediatric Surgery Department, Hospital Sant Joan de Déu, Universitat de Barcelona, Passeig Sant Joan de Déu 2, Esplugues de Llobregat, 08950, Barcelona, Spain.
authors.forename: Xavier
authors.initials: X
authors.lastname: Tarrado Castellarnau
mesh_terms: D000110:Acetylcholinesterase; D000367:Age Factors; D015415:Biomarkers / Q000378:metabolism; D001706:Biopsy; D064032:Calbindin 2 / Q000378:metabolism*; D005260:Female; D006627:Hirschsprung Disease / Q000175:diagnosis*; D006801:Humans; D007150:Immunohistochemistry; D007223:Infant; D007231:Infant, Newborn; D008297:Male; D012007:Rectum / Q000378:metabolism* / Q000473:pathology*; D012189:Retrospective Studies; D012680:Sensitivity and Specificity; D013194:Staining and Labeling; D013396:Suction
chemical_list: D015415:Biomarkers; D064032:Calbindin 2; D000110:Acetylcholinesterase
keywords: Calretinin; Hirschsprung disease; Rectal suction biopsy