Amal Dafar1, Paula Rico1, Ayşegül Işık1, Mats Jontell1, Hülya Cevik-Aras2. 1. Department of Oral Medicine and Pathology, University of Gothenburg, Odontology Institute at Sahlgrenska Academy, Gothenburg, Sweden. 2. Department of Oral Medicine and Pathology, University of Gothenburg, Odontology Institute at Sahlgrenska Academy, Gothenburg, Sweden. Electronic address: hulya.cevik@odontologi.gu.se.
Abstract
OBJECTIVES: This study aims to create consensus concerning the use of a methodology by which the handling of saliva is standardized and quantitative detection of IL-8 and EGF in whole saliva is achieved. Our study involves evaluating the extent to which the pre-treatment of saliva samples with an anionic detergent - sodium dodecyl sulphate (SDS) - improved detection levels for IL-8 and EGF. METHODS: Whole saliva samples (n=28) were collected from healthy individuals and a protease inhibitor cocktail was added immediately. They were treated with either SDS or PBS for 20min and were then applied to a sandwich ELISA. RESULTS AND CONCLUSIONS: Saliva is a complex viscous fluid that requires degrading before the analysis of salivary biomarkers. We found that pre-treatment of samples with SDS significantly increased the detection levels for both EGF (293%) and IL-8 (346%) when compared with PBS-treated pairs (***P<0.001). According to the results we recommend: (i) pre-treatment of whole saliva samples with SDS for quantitative analysis (ii) using secretory output instead of concentration in the presentation of results to avoid individual variations and (iii) taking into consideration gender, age and meal intake since these have an impact on the secretory output of salivary proteins.
OBJECTIVES: This study aims to create consensus concerning the use of a methodology by which the handling of saliva is standardized and quantitative detection of IL-8 and EGF in whole saliva is achieved. Our study involves evaluating the extent to which the pre-treatment of saliva samples with an anionic detergent - sodium dodecyl sulphate (SDS) - improved detection levels for IL-8 and EGF. METHODS: Whole saliva samples (n=28) were collected from healthy individuals and a protease inhibitor cocktail was added immediately. They were treated with either SDS or PBS for 20min and were then applied to a sandwich ELISA. RESULTS AND CONCLUSIONS: Saliva is a complex viscous fluid that requires degrading before the analysis of salivary biomarkers. We found that pre-treatment of samples with SDS significantly increased the detection levels for both EGF (293%) and IL-8 (346%) when compared with PBS-treated pairs (***P<0.001). According to the results we recommend: (i) pre-treatment of whole saliva samples with SDS for quantitative analysis (ii) using secretory output instead of concentration in the presentation of results to avoid individual variations and (iii) taking into consideration gender, age and meal intake since these have an impact on the secretory output of salivary proteins.
Authors: María José Soto-Méndez; María Eugenia Romero-Abal; Concepción María Aguilera; María Cruz Rico; Noel W Solomons; Klaus Schümann; Angel Gil Journal: PLoS One Date: 2015-06-15 Impact factor: 3.240