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Optimized RNA extraction strategy from whole human saliva
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Resumo(s)
Saliva has emerged as a potential source of biomarkers to predict, diagnosis and monitor several diseases particularly within the oral cavity (Melguizo Rodríguez et al., 2020). Using saliva to detect and quantify biomarkers offers numerous benefits over blood collection because sample is collected by a noninvasive and painless method, allowing the screening of biomarkers even in children. Moreover, saliva can be used for biomarker quantification in the context of clinical diagnosis, prognosis and disease and/or treatment monitoring of oral and systemic diseases (Song et al., 2023; Dongiovanni et al., 2023). However, saliva is a complex biofluid composed by DNA, proteins, RNA, metabolites among others, imposing experimental challenges when isolating nucleic acids, mainly RNA. This work reports different protocols to extract RNA from human saliva with and without a stabilization solution (RNAlater®, Thermo Fisher Scientific) to evaluate the best RNA extraction method for downstream gene expression studies by real-time PCR (RT-PCR). For this purpose, several parameters as RNA quality, integrity, and yield were assessed .