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Clinical Chemistry 52: 988-994, 2006. First published April 6, 2006; 10.1373/clinchem.2005.063206
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(Clinical Chemistry. 2006;52:988-994.)
© 2006 American Association for Clinical Chemistry, Inc.

Molecular Diagnostics and Genetics

Characterization of RNA in Saliva

Noh Jin Park1, Yang Li1,3, Tianwei Yu1, Brigitta M.N. Brinkman1 and David T. Wong1,2,3,4,5,a

1 Dental Research Institute,2 UCLA School of Dentistry,3 Johnson Comprehensive Cancer Center,4 Division of Head and Neck Surgery/Otolaryngology, and5 Henry Samueli School of Engineering, UCLA, Los Angeles, CA.

aAddress correspondence to this author at: UCLA School of Dentistry, Dental Research Institute, 73-017 CHS, 10833 Le Conte Ave., Los Angeles, CA 90095. Fax 310-825-7609; e-mail dtww{at}ucla.edu.

Background: We have previously shown that human mRNAs are present in saliva and can be used as biomarkers of oral cancer. In this study, we analyzed the integrity, sources, and stability of salivary RNA.

Methods: We measured the integrity of salivary RNA with reverse transcription followed by PCR (RT-PCR) or RT-quantitative PCR (RT-qPCR). To study RNA entry sites into the oral cavity, we used RT-PCR analysis of salivary RNA from the 3 major salivary glands, gingival crevice fluid, and desquamated oral epithelial cells. We measured stability of the salivary ß-actin mRNA by RT-qPCR of salivary RNA incubated at room temperature for different periods of time. We measured RNA association with other macromolecules by filtering saliva through pores of different sizes before performing RT-qPCR. To assess RNA–macromolecule interaction, we incubated saliva with Triton X-100 for different periods of time before performing RT-qPCR.

Results: In most cases, we detected partial- to full-length salivary mRNAs and smaller amounts of middle and 3' gene amplicons compared with the 5'. RNA was present in all oral fluids examined. Endogenous salivary ß-actin mRNA degraded more slowly than exogenous ß-actin mRNA, with half-lives of 12.2 and 0.4 min, respectively (P <0.001). Salivary RNA could not pass through 0.22 or 0.45 µm pores. Incubation of saliva with Triton X-100 accelerated degradation of salivary RNA.

Conclusions: Saliva harbors both full-length and partially degraded forms of mRNA. RNA enters the oral cavity from different sources, and association with macromolecules may protect salivary RNA from degradation.




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