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  • Original Article
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DNA yield and quality of saliva samples and suitability for large-scale epidemiological studies in children

International Journal of Obesity volume 35, pages S113–S118 (2011)Cite this article

Subjects

Abstract

Objective:

To evaluate two saliva collection methods for DNA yield and quality as applied to a large, integrated, multicentre, European project involving the collection of biological material from children.

Design:

Cross-sectional multicentre comparative study in young children.

Methods:

Saliva samples were collected from 14 019 children aged 2–9 years from eight European countries participating in the IDEFICS (Identification and prevention of dietary- and lifestyle-induced health effects in children and infants) study. This involved either the collection of 2 ml of saliva from children who were able to spit, or using a sponge to collect whole saliva and buccal mucosal cells from the inside of the mouth of younger children unable to spit. Samples were assembled centrally in each participating centre and subsequently despatched for DNA extraction and biobanking to the University of Glasgow. A subgroup of 4678 samples (∼33% of sampled individuals) were chosen for DNA extraction before genotyping.

Results:

The whole-saliva collection method resulted in a higher DNA yield than the sponge collection method (mean±s.d.; saliva: 20.95±2.35 μg, sponge: 9.13±2.25 μg; P<0.001). DNA quality as measured by A260/A280 was similar for the two collection methods. A minimum genotype calling success rate of 95% showed that both methods provide good-quality DNA for genotyping using TaqMan allelic discrimination assays.

Conclusions:

Our results showed higher DNA yield from the whole-saliva collection method compared with the assisted sponge collection. However, both collection methods provided DNA of sufficient quantity and quality for large-scale genetic epidemiological studies.

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Acknowledgements

We acknowledge all members of the UGLW DNA extraction team for their excellent technical assistance in completing the DNA extraction process. This study was conducted as part of the IDEFICS study (http://www.idefics.eu). We gratefully acknowledge the financial support of the European Community within the Sixth RTD Framework Programme Contract No. 016181 (FOOD).

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Authors and Affiliations

  1. College of Medicine, Veterinary and Life Sciences, Institute of Cardiovascular and Medical Sciences, University of Glasgow, Glasgow, Scotland, UK

    A C Koni, R A Scott, G Wang & Y P Pitsiladis

  2. School of Life Sciences, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow, Scotland, UK

    M E S Bailey

  3. Division of Epidemiological Methods and Etiologic Research, Bremen Institute for Prevention Research and Social Medicine, University of Bremen, Bremen, Germany

    J Peplies

  4. Department of Biometry, Bremen Institute for Prevention Research and Social Medicine, University of Bremen, Bremen, Germany

    K Bammann

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  1. A C Koni
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Correspondence to Y P Pitsiladis.

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The authors declare no conflict of interest.

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Statement of ethics

We certify that all applicable institutional and governmental regulations pertaining to the ethical use of human volunteers were followed during this research. Approval by the appropriate ethics committees was obtained by each of the eight centres carrying out the fieldwork. Participants were not subjected to any study procedure before both the children and their parents gave consent for examinations, collection of samples, subsequent analysis and storage of personal data and collected samples. The participating children and their parents could consent to single components of the study while refraining from others.

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Koni, A., Scott, R., Wang, G. et al. DNA yield and quality of saliva samples and suitability for large-scale epidemiological studies in children. Int J Obes 35 (Suppl 1), S113–S118 (2011). https://doi.org/10.1038/ijo.2011.43

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