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Salivary Diagnostics Using Purified Nucleic Acids

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Book cover Oral Biology

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1537))

Abstract

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.

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References

  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: 978-1-4557-3127-5

    Google Scholar 

  2. Streckfus C, Bigler L (2002) Saliva as a diagnostic fluid. Oral Dis 8:69–76

    Article  CAS  PubMed  Google Scholar 

  3. Slowey PD (2013) Commercial saliva collections tools. J Calif Dent 41(2):97–105

    Google Scholar 

  4. Malamud D, Tabak LA (eds) (1992) Saliva as a diagnostic fluid. Vol 694, Annals NY Academy of Science

    Google Scholar 

  5. Samaranayake LP (2006) Saliva tells the body’s health. In: Stephen Moss (ed) The benefits of chewing. pp 24–35

    Google Scholar 

  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):183

    Google Scholar 

  7. Testing Oral Fluid for the Presence of HIV Antibodies February 2013. Report by the Association of Public Health Laboratories (APHL) http://www.aphl.org/AboutAPHL/publications/Documents/ID_Feb2013_Testing-of-Oral-Fluid-for-the-Presence-of-HIV-Antibodies-Brief.pdf

  8. See http://www.zrtlab.com/patients-standard-tests/saliva

  9. See http://www.diagnostechs.com/Pages/Home.aspx

  10. See https://www.labrix.com/SalivaryHormoneTesting

  11. Cone EJ, Huestis M (2007) Interpretation of oral fluid tests for drugs of abuse. Ann NY Acad Sci 1098:51–103

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Bradshaw DJ, Marsh PD (1998) Analysis of pH-driven disruption of oral microbial communities in vitro. Caries Res 32:456–462

    Article  CAS  PubMed  Google Scholar 

  13. Bratthall D, Hansel Petersson G (2005) Cariogram-a multifactorial risk assessment model for a multifactorial disease. Community Dent Oral Epidemiol 33:256–264

    Article  PubMed  Google Scholar 

  14. Larmas M (1992) Saliva and dental caries: diagnostic tests for normal dental practice. Int Dent J 42:199–208

    CAS  PubMed  Google Scholar 

  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 1098:411–428

    Article  CAS  PubMed  Google Scholar 

  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 10:8442–8450

    Article  CAS  PubMed  Google Scholar 

  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 18:17–24

    Article  CAS  PubMed  Google Scholar 

  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 91:174–179

    Article  CAS  PubMed  Google Scholar 

  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 6:2363–2370

    CAS  PubMed  Google Scholar 

  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 3:138–143

    Article  CAS  PubMed  Google Scholar 

  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 18:101–109

    Article  CAS  PubMed  Google Scholar 

  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 33:595–600

    Article  CAS  PubMed  Google Scholar 

  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 5:321–326

    Article  Google Scholar 

  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 60:81–92

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Corstjens LAM, Abrams WR, Malamud D (2012) Detecting viruses by using salivary diagnostics. J Am Dent Assoc 143:12S–18S

    Article  PubMed  PubMed Central  Google Scholar 

  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 37:505–514

    Google Scholar 

  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, http://www.Plosone.org

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. See http://www.dnagenotek.com/US/pdf/MK-006.pdf

  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 2014

    Google Scholar 

  30. See https://norgenbiotek.com/product/saliva-dna-collection-preservation-and-isolation-kit

  31. See http://biomatrica.com/dnagardsaliva.php

  32. See http://www.mawidna.com/products/iswab-dna-collection-kit

  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 2014

    Google Scholar 

  34. See http://www.isohelix.com/products/isohelix-dna-buccal-swabs

  35. Lee YH, Zhou H, Yan X, Zhang L, Chia D, Wong DTW (2011) Direct saliva transcriptome analysis. Clin Chem 57:1295–1302

    Article  CAS  PubMed  Google Scholar 

  36. See http://www.dnagenotek.com/US/products/RE100.html

  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 56:1506–1513

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. See https://norgenbiotek.com/product/saliva-rna-collection-and-preservation-devices

  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 58:69–76

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  40. See http://4saliva.com/products/pure%E2%80%A2sal

  41. Théry C, Zitvogel L, Amigorena S (2002) Exosomes: composition, biogenesis and function. Nat Rev Immunol 2:569–579

    PubMed  Google Scholar 

  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 16:34–38

    Article  CAS  PubMed  Google Scholar 

  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 288:26888–26897

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  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 9:9

    Article  Google Scholar 

  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 2014

    Google Scholar 

  46. See https://www.systembio.com/microrna-research/exoquick-exosomes/overview?gclid=CPTmjv2foMoCFQWUfgodu_gJUw

  47. Savina A, Vidal M, Colombo MI (2002) The exosome pathway in K562 cells is regulated by Rab11. J Cell Sci 115:2505–2515

    CAS  PubMed  Google Scholar 

  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–H3056

    Article  CAS  PubMed  Google Scholar 

  49. Fleischhacker M, Schmidt B (2007) Circulating nucleic acids (CNAs) and cancer—a survey. Biochim Biophys Acta 1775:181–232

    CAS  PubMed  Google Scholar 

  50. The American College of Obstetricians and Gynecologists Committee Opinion Number 640 September 2015. Cell-free DNA screening for fetal aneuploidy, http://www.acog.org/Resources-And-Publications/Committee-Opinions/Committee-on-Genetics/Cell-free-DNA-Screening-for-Fetal-Aneuploidy

  51. Zhong XY, von Mühlenen I, Li Y (2007) Increased concentrations of antibody-bound circulatory cell-free DNA in rheumatoid arthritis. Clin Chem 53:1609–1614

    Article  CAS  PubMed  Google Scholar 

  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 49:1286–1291

    Article  CAS  PubMed  Google Scholar 

  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 327:95–101

    Article  CAS  PubMed  Google Scholar 

  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 47:253–258

    Article  CAS  PubMed  Google Scholar 

  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 2014

    Google Scholar 

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Acknowledgments

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

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Correspondence to Paul D. Slowey .

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Slowey, P.D. (2017). Salivary Diagnostics Using Purified Nucleic Acids. In: Seymour, G., Cullinan, M., Heng, N. (eds) Oral Biology. Methods in Molecular Biology, vol 1537. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6685-1_1

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  • DOI: https://doi.org/10.1007/978-1-4939-6685-1_1

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6683-7

  • Online ISBN: 978-1-4939-6685-1

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