Salivary proteomic profile of babies with Congenital Syndrome associated with Zika Virus

• Grant number: 16/23913-0
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: July 01, 2017
• End date: June 30, 2018
• Field of knowledge: Health Sciences - Dentistry - Pediatric Dentistry
• Principal Investigator: Maria Aparecida de Andrade Moreira Machado
• Grantee: Adassa Bianca Gomes Silva
• Host Institution: Faculdade de Odontologia de Bauru (FOB). Universidade de São Paulo (USP). Bauru , SP, Brazil

Abstract

The use of saliva is an option that should be considered for health and disease monitoring parameters not just because of its various components, but also and this is a non-invasive, painless and easily obtainable method. Identifying and measuring salivary biomarkers creates perspectives for the diagnosis, including early detection of disease and monitoring its progression, anticipating the initiation of treatment. The present study aims to explore the use of saliva for the detection of RNA of the Zika virus (ZIKAV) in infants with syndrome associated with congenital ZIKAV. The sample will be composed of 40 babies, 20 healthy, for negative control, and 20 which present congenital syndrome associated with Zika virus (microcephaly). The mothers will respond to a questionnaire, reporting the signs and symptoms of the disease during pregnancy and how this disease was diagnosed. Saliva will be colleted using Siqueira's methodology (Siqueira et al., 2005). In order to avoid the degradation of total salivary RNA, the RNAlater® stabilizing solution (Sigma-Aldrich) will be added to the samples. Samples will be frozen at -80 °C until further analysis. Initially, the total RNA extractions from the saliva samples will be performed using the QIAamp Viral RNA Mini Kit (52906-Applied Biosystems, Life Technologies, USA), followed by the quantification and evaluation of RNA quality through the spectrophotometer. Then, the treatment of all samples of total RNA with DNAse will be performed in order to avoid the possibility of contamination by genomic DNA. Posteriorly, reverse transcription will be performed followed by Quantitative Polymerase Chain Reaction (RT-qPCR). The results will be analyzed based on the Ct values (cicle threshold), which will be used as the point corresponding to the number of cycles where the amplification reaches a threshold, which allows the quantitative analysis of the target gene expression, and then will be compared the differences between the cycles of the samples from patient with and without syndrome associated with congenital Zika virus (microcephaly). The data obtained will be analyzed by means of the Analysis of Variance (ANOVA) test for one criterion. The test between groups will be applied the Tukey Test for multiple comparisons, adopting a level of significance of 5%. (AU)