title: Reproducibility, repeatability, and level of difficulty of two methods for tumor budding evaluation in oral squamous cell carcinoma.
abstract: BACKGROUND
  This study aimed to analyze the reproducibility, repeatability, and level of difficulty of two methods for tumor budding evaluation in oral squamous cell carcinoma (OSCC): staining by hematoxylin and eosin (HE) and immunostaining for multicytokeratin.
  
  
  METHODS
  The evaluation of tumor budding was performed by three examiners in 103 samples of OSCC, using the two methods. A Likert-type scale was used to measure the difficulty in the assessment. The interexaminer agreement (reproducibility) was estimated using Fleiss's kappa and the intra-examiner agreement (repeatability) was estimated using Cohen's kappa. The agreement between the two methods was evaluated using Cohen's Kappa. The Friedman test was used to compare the three examiners' perceived levels of difficulty of assessment. The Wilcoxon test was used to compare the level of difficulty of the evaluation between the two methods.
  
  
  RESULTS
  Reproducibility by the immunostaining method for multicytokeratin was substantial, being higher than the only fair agreement by the HE. Repeatability by the HE ranged from moderate to substantial among examiners, regardless of the examiner's experience. Repeatability by the immunostaining method for multicytokeratin did not vary among examiners, showing almost perfect agreement. The agreement between the two methods ranged from fair to moderate among examiners, being lower in the less experienced examiner. All the examiners presented greater difficulty in the evaluation by the HE.
  
  
  CONCLUSIONS
  In view of the unsatisfactory agreement between the two methods of tumor budding evaluation in OSCC, it is recommended that this evaluation should be performed by the immunostaining method for multicytokeratin, considering its higher reproducibility, greater replicability, and lower difficulty compared to the HE.
authors.affiliation: Oral Pathology Section, School of Dentistry, Pontifical Catholic University of Minas Gerais (PUC Minas), Belo Horizonte, MG, Brazil.
authors.forename: Priscila Laiza Rubim
authors.initials: PLR
authors.lastname: Leão
authors.affiliation: Oral Pathology Section, School of Dentistry, Pontifical Catholic University of Minas Gerais (PUC Minas), Belo Horizonte, MG, Brazil.
authors.forename: Helvécio
authors.initials: H
authors.lastname: Marangon Junior
authors.affiliation: Oral Pathology Section, School of Dentistry, Pontifical Catholic University of Minas Gerais (PUC Minas), Belo Horizonte, MG, Brazil.
authors.forename: Victória Vasconcellos Moreira
authors.initials: VVM
authors.lastname: Melo
authors.affiliation: Oral Pathology Section, School of Dentistry, Pontifical Catholic University of Minas Gerais (PUC Minas), Belo Horizonte, MG, Brazil.
authors.forename: Ângela Braga
authors.initials: ÂB
authors.lastname: Caixeta
authors.affiliation: Oral Pathology Section, School of Dentistry, Pontifical Catholic University of Minas Gerais (PUC Minas), Belo Horizonte, MG, Brazil.
authors.forename: Paulo Eduardo Alencar
authors.initials: PEA
authors.lastname: Souza
authors.affiliation: Department of Oral Diagnosis and Pathology, School of Dentistry, Federal University of Minas Gerais (UFMG), Belo Horizonte, MG, Brazil.
authors.forename: Maria Cássia Ferreira
authors.initials: MCF
authors.lastname: de Aguiar
authors.affiliation: Oral Pathology Section, School of Dentistry, Pontifical Catholic University of Minas Gerais (PUC Minas), Belo Horizonte, MG, Brazil.
authors.forename: Martinho Campolina Rebello
authors.identifier: http://orcid.org/0000-0003-0192-5614
authors.initials: MCR
authors.lastname: Horta
mesh_terms: D002294:Carcinoma, Squamous Cell / Q000473:pathology*; D004396:Coloring Agents; D004801:Eosine Yellowish-(YS); D005260:Female; D005456:Fluorescent Dyes; D006416:Hematoxylin; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods*; D008297:Male; D008875:Middle Aged; D009062:Mouth Neoplasms / Q000473:pathology*; D010338:Pathology, Clinical / Q000379:methods; D015203:Reproducibility of Results; D013194:Staining and Labeling / Q000379:methods*
chemical_list: D004396:Coloring Agents; D005456:Fluorescent Dyes; D004801:Eosine Yellowish-(YS); D006416:Hematoxylin
keywords: oral cancer; oral squamous cell carcinoma; tumor budding