Skip to main content
Key Specialties
Cardiology Diabetology Endocrinology Gastroenterology Geriatrics and Gerontology Gynecology and Obstetrics Hematology Infectious Disease Internal Medicine Nephrology Neurology Oncology Pediatrics Respiratory Medicine Rheumatology Surgery
Congress Coverage
2024 ACC Congress 2023 ESMO Congress 2023 EASD Congress 2023 ERS Congress 2023 ESC Congress
Clinical Collections
Advances in Alzheimer’s

At a glance: The STEP trials

A round-up of the STEP phase 3 clinical trials evaluating semaglutide for weight loss in people with overweight or obesity.
ADVANCED SEARCH
Log in
Top
Clinical Oral Investigations
Published in: Clinical Oral Investigations 7/2013

01-09-2013 | Original Article

Evaluation of the long-term storage stability of saliva as a source of human DNA

Authors: Robert P. Anthonappa, Nigel M. King, A. Bakr M. Rabie

Published in: Clinical Oral Investigations | Issue 7/2013

Login to get access

Abstract

Objectives

The objectives of this paper are to determine the storage stability of saliva at 37 °C over an 18-month period, and its influence on the DNA yield, purity, PCR protocols and genotyping efficacy.

Materials and methods

Of the 60 participants, blood samples were obtained from 10 and saliva from 50. Samples were subjected to different storage conditions: DNA extracted immediately; DNA extracted following storage at 37 °C for 1, 6, 12 and 18 months. Subsequently, DNA yield, OD260/280 and OD260/230 ratios were measured. The isolated DNA was used to amplify exons 0–7 of the RUNX2 gene and subsequently sequenced. Furthermore, 25 SNPs were genotyped.

Results

The mean DNA yield, OD260/280 and OD260/230 ratios obtained from blood were 67.4 ng/μl, 1.8 ± 0.05 and 1.8 ± 0.4 respectively. DNA yield obtained from saliva was significantly higher than blood (p < 0.0001), ranging from 97.4 to 125.8 ng/μl while the OD260/280 ratio ranged from 1.8 ± 0.13 to 1.9 ± 0.1. The success rates for the 25 SNPs ranged from 98 to 100 % for blood and 96–99 % for saliva samples with the genotype frequencies in Hardy–Weinberg equilibrium (>0.01).

Conclusions

Saliva can be stored at 37 °C for 18 months without compromising its quality and ability to endure genetic analyses.

Clinical relevance

Saliva is a viable source of human DNA to facilitate the feasibility of large-scale genetic studies.
Literature
1.
Ng DP, Koh D, Choo S, Chia KS (2006) Saliva as a viable alternative source of human genomic DNA in genetic epidemiology. Clin Chim Acta 367:81–85PubMedCrossRef
2.
Ng DP, Koh D, Choo S, Ng V, Fu QY (2004) Effect of storage conditions on the extraction of PCR-quality genomic DNA from saliva. Clin Chim Acta 343:191–194PubMedCrossRef
3.
Quinque D, Kittler R, Kayser M, Stoneking M, Nasidze I (2006) Evaluation of saliva as a source of human DNA for population and association studies. Anal Biochem 353:272–277PubMedCrossRef
4.
Rogers NL, Cole SA, Lan HC, Crossa A, Demerath EW (2007) New saliva DNA collection method compared to buccal cell collection techniques for epidemiological studies. Am J Hum Biol 19:319–326PubMedCrossRef
5.
Mulot C, Stücker I, Clavel J, Beaune P, Loriot MA (2005) Collection of human genomic DNA from buccal cells for genetics studies: comparison between cytobrush, mouthwash, and treated card. J Biomed Biotechnol 3:291–296CrossRef
6.
García-Closas M, Egan KM, Abruzzo J, Newcomb PA, Titus-Ernstoff L, Franklin T et al (2001) Collection of genomic DNA from adults in epidemiological studies by buccal cytobrush and mouthwash. Cancer Epidemiol Biomarkers Prev 6:687–696
7.
Cozier YC, Palmer JR, Rosenberg L (2004) Comparison of methods for collection of DNA samples by mail in the Black Women’s Health Study. Ann Epidemiol 2:117–122CrossRef
8.
Luan JA, Wong MY, Day NE, Wareham NJ (2001) Sample size determination for studies of gene–environment interaction. Int J Epidemiol 30:1035–1040PubMedCrossRef
9.
Holland NT, Smith MT, Eskenazi B, Bastaki M (2003) Biological sample collection and processing for molecular epidemiological studies. Mutat Res 543:217–234PubMedCrossRef
10.
Xuan D, Li S, Zhang X, Hu F, Lin L, Wang C et al (2008) Mutations in the RUNX2 gene in Chinese patients with cleidocranial dysplasia. Ann Clin Lab Sci 38:15–24PubMed
11.
Phillips MS, Lawrence R, Sachidanandam R, Morris AP, Balding DJ, Donaldson MA et al (2003) Chromosome-wide distribution of haplotype blocks and the role of recombination hot spots. Nat Genet 33:382–387PubMedCrossRef
12.
Halsall A, Ravetto P, Reyes Y, Thelwell N, Davidson A, Gaut R et al (2008) The quality of DNA extracted from liquid or dried blood is not adversely affected by storage at 4 degrees C for up to 24 h. Int J Epidemiol 37(Suppl 1):i7–i10PubMedCrossRef
13.
Visvikis S, Schlenck A, Maurice M (1998) DNA extraction and stability of epidemiological studies. Clin Chem Lab Med 36:551–555PubMedCrossRef
14.
Dawes C (2003) Estimates, from salivary analyses, of the turnover time of the oral mucosal epithelium in humans and the number of bacteria in an edentulous mouth. Arch Oral Biol 48:329–336PubMedCrossRef
15.
Halsall A, Ravetto P, Reyes Y, Thelwell N, Davidson A, Gaut R et al (2008) The quality of DNA extracted from liquid or dried blood is not adversely affected by storage at 4 degrees C for up to 24 h. Int J Epidemiol 37:7–10CrossRef
16.
Rylander-Rudqvist T, Håkansson N, Tybring G, Wolk A (2006) Quality and quantity of saliva DNA obtained from the self-administrated oragene method—a pilot study on the cohort of Swedish men. Cancer Epidemiol Biomarkers Prev 15:1742–1745PubMedCrossRef
17.
Chartier J, Birnboim H (2004) Bacterial DNA content with OrageneTM. DNA Genotek, Ottawa
18.
van Schie RC, Wilson ME (1997) Saliva: a convenient source of DNA for analysis of bi-allelic polymorphisms of Fc gamma receptor IIA (CD32) and Fc gamma receptor IIIB (CD16). J Immunol Methods 208:91–101PubMedCrossRef
19.
Andrisin TE, Humma LM, Johnson JA (2002) Collection of genomic DNA by the non-invasive mouthwash method for use in pharmacogenetic studies. Pharmacotherapy 22:954–960PubMedCrossRef
Metadata
Title
Evaluation of the long-term storage stability of saliva as a source of human DNA
Authors
Robert P. Anthonappa
Nigel M. King
A. Bakr M. Rabie
Publication date
01-09-2013
Publisher
Springer Berlin Heidelberg
Published in
Clinical Oral Investigations / Issue 7/2013
Print ISSN: 1432-6981
Electronic ISSN: 1436-3771
DOI
https://doi.org/10.1007/s00784-012-0871-5

Other articles of this Issue 7/2013

Clinical Oral Investigations 7/2013 Go to the issue

Letter to the Editor

Comment on “Development of digital shade guides for color assessment using a digital camera with ring flashes [Oi-Hong Tung et al., Clinical Oral Investigations (2011) 15:49–56]”

Letter to the Editor

More about accuracy of peri-implant bone thickness and validity of assessing bone augmentation material using cone beam computed tomography

Original Article

Clinical and microbiological changes after minimally invasive therapeutic approaches in intrabony defects: a 12-month follow-up

Erratum

Erratum to: A micro-computed tomography study of canal configuration of multiple-canalled mesiobuccal root of maxillary first molar

Original Article

Root resorption of endodontically treated teeth following orthodontic treatment: a meta-analysis

Original Article

Role of amniotic fluid mesenchymal cells engineered on MgHA/collagen-based scaffold allotransplanted on an experimental animal study of sinus augmentation