Journal
ARCHIVES OF ORAL BIOLOGY
Volume 54, Issue 3, Pages 268-273Publisher
PERGAMON-ELSEVIER SCIENCE LTD
DOI: 10.1016/j.archoralbio.2008.10.004
Keywords
Saliva diagnostics; Saliva stabilizer; Salivary biomarkers; Proteins; DNA; RNA
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Funding
- [R21 CA126733 U01-DE017790]
- [R03-DE017144]
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Objective: Saliva is a biofluid that can be obtained from individuals without supervision by health care providers. To maximize this clinical advantage, it is highly desirable to have a global salivary analyte stabilizer for proteins, RNA and DNA at ambient temperature. Design: Whole saliva, saliva supernatant and saliva filtrate (5.0 mu m) were treated with RPS at room temperature (RT) for up to 6 days and then subjected to SDS-PAGE. Immunoblotting of beta-actin and cystatin C were used to evaluate protein stability. For salivary DNA/RNA, whole saliva was incubated with RPS at RT for up to 10 weeks. After extracting total DNA/RNA in samples at week 0, 2,6 and 10, DNA stability was assayed by chromosome 18 DNA qPCR and RNA stability by beta-actin mRNA RT-qPCR. Results: beta-actin completely degraded in all types of saliva samples after 6-day incubation at RT. However, 24.0%, 91.4% and 89.3% of beta-actin remained intact with RPS for whole saliva, saliva supernatant and filtrate, respectively. Similarly, 70.3% of cystatin C in supernatant remained intact in the presence of RPS. For salivary DNA/RNA, the cycle threshold (Ct) values showed no significant change for chromosome 18 DNA and beta-actin mRNA in RPS-incubated saliva during the 10-week time course while significant increase in Ct values were observed in controls without RPS for both beta-actin mRNA and DNA. Conclusions: RPS provided effective concurrent stabilization to salivary DNA/RNA in whole saliva for up to 10 weeks and proteins in saliva filtrate for 6 days at RT. We also achieved separation of saliva supernatant from cellular elements by a simple filtration step (bypassing the need for centrifugation). (c) 2008 Elsevier Ltd. All rights reserved.
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