Salivary Diagnostics Using Purified Nucleic Acids

  • Paul D. SloweyEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1537)


Saliva is an easily accessible fluid that has led to increasing interest in the development of salivary diagnostics. This chapter describes some of the newer tools and procedures for collection, stabilization, and storage of oral fluid matrices that aid in the successful use of saliva as a test specimen. This chapter focuses particularly on nucleic acid components for downstream molecular diagnostic (MDx) testing, since this is probably the area where saliva is likely to have the greatest impact in improving healthcare for the general population.

Key words

Saliva RNA DNA Nucleic acids Stabilization Exosomes 



The author would like to acknowledge the support of Dr David T Wong (UCLA) for his support and encouragement in preparing this manuscript.


  1. 1.
    Punyadeera C, Slowey PD (2013) Saliva as an emerging biofluid for clinical diagnosis and applications of MEMS/NEMS in salivary diagnostics. Chapter 22 in “Nanobiomaterials in Clinical Dentistry.” Elsevier, pp 453–475, ISBN: 978-1-4557-3127-5Google Scholar
  2. 2.
    Streckfus C, Bigler L (2002) Saliva as a diagnostic fluid. Oral Dis 8:69–76CrossRefPubMedGoogle Scholar
  3. 3.
    Slowey PD (2013) Commercial saliva collections tools. J Calif Dent 41(2):97–105Google Scholar
  4. 4.
    Malamud D, Tabak LA (eds) (1992) Saliva as a diagnostic fluid. Vol 694, Annals NY Academy of ScienceGoogle Scholar
  5. 5.
    Samaranayake LP (2006) Saliva tells the body’s health. In: Stephen Moss (ed) The benefits of chewing. pp 24–35Google Scholar
  6. 6.
    Zachary D, Mwenge L, Muyoyeta M, Shanaube K, Schaap A, Bond V, Kosloff B, de Haas P, Ayles HBMC (2012) Infect Dis 8(12):183Google Scholar
  7. 7.
    Testing Oral Fluid for the Presence of HIV Antibodies February 2013. Report by the Association of Public Health Laboratories (APHL)
  8. 8.
  9. 9.
  10. 10.
  11. 11.
    Cone EJ, Huestis M (2007) Interpretation of oral fluid tests for drugs of abuse. Ann NY Acad Sci 1098:51–103CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Bradshaw DJ, Marsh PD (1998) Analysis of pH-driven disruption of oral microbial communities in vitro. Caries Res 32:456–462CrossRefPubMedGoogle Scholar
  13. 13.
    Bratthall D, Hansel Petersson G (2005) Cariogram-a multifactorial risk assessment model for a multifactorial disease. Community Dent Oral Epidemiol 33:256–264CrossRefPubMedGoogle Scholar
  14. 14.
    Larmas M (1992) Saliva and dental caries: diagnostic tests for normal dental practice. Int Dent J 42:199–208PubMedGoogle Scholar
  15. 15.
    Christodoulides N, Floriano PN, Miller CS, Ebersole JL, Mohanty S, Dharshan P, Griffin M, Lennart A, Ballard KL, King CP Jr, Langub MC, Kryscio RJ, Thomas MV, McDevitt JT (2007) Lab-on-a-chip methods for point-of-care measurements of salivary biomarkers of periodontitis. Ann NY Acad Sci 1098:411–428CrossRefPubMedGoogle Scholar
  16. 16.
    Li Y, St John MA, Zhou X, Kim Y, Sinha U, Jordan RC, Eisele D, Abemayor E, Elashoff D, Park NH, Wong DT (2004) Salivary transcriptome diagnostics for oral cancer detection. Clin Cancer Res 10:8442–8450CrossRefPubMedGoogle Scholar
  17. 17.
    Streckfus C, Bigler L (2005) The use of soluble, salivary c-erbB-2 for the detection and post-operative follow-up of breast cancer in women: the results of a five-year translational research study. Adv Dent Res 18:17–24CrossRefPubMedGoogle Scholar
  18. 18.
    Streckfus C, Bigler L, Dellinger T, Dai X, Cox WJ, McArthur A, Kingman A, Thigpen JT (2001) Reliability assessment of soluble c-erbB-2 concentrations in the saliva of healthy women and men. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 91:174–179CrossRefPubMedGoogle Scholar
  19. 19.
    Streckfus C, Bigler L, Dellinger T, Dai X, Kingman A, Thigpen JT (2000) The presence of soluble c-erbB-2 in saliva and serum among women with breast carcinoma: a preliminary study. Clin Cancer Res 6:2363–2370PubMedGoogle Scholar
  20. 20.
    Streckfus C, Bigler L, Dellinger T, Pfeifer M, Rose A, Thigpen JT (1999) CA 15-3 and c-erbB-2 presence in the saliva of women. Clin Oral Investig 3:138–143CrossRefPubMedGoogle Scholar
  21. 21.
    Streckfus C, Bigler L, Tucci M, Thigpen JT (2000) A preliminary study of CA15-3, c-erbB-2, epidermal growth factor receptor, cathepsin-D, and p53 in saliva among women with breast carcinoma. Cancer Invest 18:101–109CrossRefPubMedGoogle Scholar
  22. 22.
    Streckfus C, Bigler L, Dellinger T, Kuhn M, Chouinard N, Dai X (2004) The expression of the c-erbB-2 receptor protein in glandular salivary secretions. J Oral Pathol Med 33:595–600CrossRefPubMedGoogle Scholar
  23. 23.
    Ovchinnikov DA, Cooper MA, Pandit P, Coman WB, Cooper-White JJ, Keith P, Wolvetang EJ, Slowey PD, Punyadeera C (2012) Tumor-suppressor gene promoter hypermethylation in saliva of head and neck cancer patients. Translat Oncol 5:321–326CrossRefGoogle Scholar
  24. 24.
    Hu S, Zhou M, Jiang J, Wang J, Elashoff D, Gorr S, Michie SA, Spijkervet FK, Bootsma H, Kallenberg CG, Vissink A, Horvath S, Wong DT (2009) Systems biology analysis of Sjogren’s syndrome and mucosa-associated lymphoid tissue lymphoma in parotid glands. Arthritis Rheum 60:81–92CrossRefPubMedPubMedCentralGoogle Scholar
  25. 25.
    Corstjens LAM, Abrams WR, Malamud D (2012) Detecting viruses by using salivary diagnostics. J Am Dent Assoc 143:12S–18SCrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    Shirtcliff EA, Buck RL, Laughlin MJ, Hart T, Cole CR, Slowey PD (2015) Salivary cortisol results obtainable within minutes of sample collection correspond with traditional immunoassays. Clin Ther 37:505–514Google Scholar
  27. 27.
    Gallo A, Tandon M, Alevizos I, Illie GG (2012) The majority of MicroRNAs detectable in serum and saliva is concentrated in exosomes. PLoS One 7, e30679, CrossRefPubMedPubMedCentralGoogle Scholar
  28. 28.
  29. 29.
    Thomas GA, Oberkanins C, Berndt A, Slowey PD (2014) Validation of a series of genomic StripAssays® to salivary DNA collection using the DNA⋅SAL™ Device. Paper presented at the American Society of Human Genetics Meeting, San Diego, October 2014Google Scholar
  30. 30.
  31. 31.
  32. 32.
  33. 33.
    El-Hahmawi B (2014) An efficient non-invasive sample collection technology for various population segments. Paper presented at the Qatar Foundation Annual Research Conference, At Doha, Qatar, Accessed 18 Nov 2014Google Scholar
  34. 34.
  35. 35.
    Lee YH, Zhou H, Yan X, Zhang L, Chia D, Wong DTW (2011) Direct saliva transcriptome analysis. Clin Chem 57:1295–1302CrossRefPubMedGoogle Scholar
  36. 36.
  37. 37.
    Patel RS, Jakymiw A, Yao B, Pauley BA, Carcamo WC, Katz J, Cheng JQ, Chan EK (2011) High resolution of microRNA signatures in human whole saliva. Arch Oral Biol 56:1506–1513CrossRefPubMedPubMedCentralGoogle Scholar
  38. 38.
  39. 39.
    Chiang SH, Thomas GA, Liao W, Grogan T, Buck RL, Fuentes L, Yakob M, Laughlin MJ, Schafer C, Nazmul-Hossain A, Wei F, Elashoff D, Slowey PD, Wong DT (2015) RNAPro*SAL: a device for rapid and standardized collection of saliva RNA and proteins. Biotechniques 58:69–76CrossRefPubMedPubMedCentralGoogle Scholar
  40. 40.
  41. 41.
    Théry C, Zitvogel L, Amigorena S (2002) Exosomes: composition, biogenesis and function. Nat Rev Immunol 2:569–579PubMedGoogle Scholar
  42. 42.
    Michael A, Bajracharya SD, Yuen PS, Zhou H, Star RA, Illei GG, Alevizos I (2010) Exosomes from human saliva as a source of microRNA biomarkers. Oral Dis 16:34–38CrossRefPubMedGoogle Scholar
  43. 43.
    Lau C, Kim Y, Chia D, Spielmann N, Eibl G, Elashoff D, Wei F, Lin YL, Moro A, Grogan T, Chiang S, Feinstein E, Schafer C, Farrell J, Wong DT (2013) Role of pancreatic cancer-derived exosomes in salivary biomarker development. J Biol Chem 288:26888–26897CrossRefPubMedPubMedCentralGoogle Scholar
  44. 44.
    Lässer C, Alikhani VS, Ekström K, Eldh M, Paredes PT, Bossios A, Sjöstran M, Gabrielsson S, Lötvall J, Valadi H (2011) Human saliva, plasma and breast milk exosomes contain RNA: uptake by macrophages. J Translat Med 9:9CrossRefGoogle Scholar
  45. 45.
    Laughlin MJ, Buck RL, Slowey PD (2014) A simplified method for the isolation of exosomes from saliva using a prototype saliva collection device (RPSAL-702). Paper presented at the North American Saliva Symposium Boston, October 2014Google Scholar
  46. 46.
  47. 47.
    Savina A, Vidal M, Colombo MI (2002) The exosome pathway in K562 cells is regulated by Rab11. J Cell Sci 115:2505–2515PubMedGoogle Scholar
  48. 48.
    Gupta S, Knowlton AA (2007) HSP60 trafficking in adult cardiac myocytes: role of the exosomal pathway. Am J Physiol Heart Circ Physiol 292:H3052–H3056CrossRefPubMedGoogle Scholar
  49. 49.
    Fleischhacker M, Schmidt B (2007) Circulating nucleic acids (CNAs) and cancer—a survey. Biochim Biophys Acta 1775:181–232PubMedGoogle Scholar
  50. 50.
    The American College of Obstetricians and Gynecologists Committee Opinion Number 640 September 2015. Cell-free DNA screening for fetal aneuploidy,
  51. 51.
    Zhong XY, von Mühlenen I, Li Y (2007) Increased concentrations of antibody-bound circulatory cell-free DNA in rheumatoid arthritis. Clin Chem 53:1609–1614CrossRefPubMedGoogle Scholar
  52. 52.
    Lam NY, Rainer TH, Chan LY, Joynt GM, Lo YM (2003) Time course of early and late changes in plasma DNA in trauma patients. Clin Chem 49:1286–1291CrossRefPubMedGoogle Scholar
  53. 53.
    Chang CP, Chia RH, Wu TL, Tsao KC, Sun CF, Wu JT (2003) Elevated cell-free serum DNA detected in patients with myocardial infarction. Clin Chim Acta 327:95–101CrossRefPubMedGoogle Scholar
  54. 54.
    Moreira VG, Prieto B, Rodriguez JS, Alvarez FV (2010) Usefulness of cell free plasma DNA, procalcitonin and C-reactive protein as markers of infection in febrile patients. Ann Clin Biochem 47:253–258CrossRefPubMedGoogle Scholar
  55. 55.
    Slowey PD, Giese U, Hofner M, Kegler U, Weber M, Buck RL, Laughlin MJ (2014) Comparison of RNAPro⋅SAL™ saliva collection versus centrifugation for cell-free DNA isolation from saliva specimens. Paper presented at the Molecular Medicine Tri-Conference, San Francisco CA, February 2014Google Scholar

Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  1. 1.Oasis Diagnostics® CorporationVancouverUSA

Personalised recommendations