DNA quality from buccal swabs in neonates: comparison of different storage time

  • Klara Yuliarti Doctoral Program in Medicine, Faculty of Medicine, Universitas Indonesia, Jakarta, Indonesia; Department of Pediatrics, Faculty of Medicine, Universitas Indonesia, Cipto Mangunkusumo Hospital, Jakarta, Indonesia; Human Genetic Research Center, Indonesian Medical Education and Research Institute (IMERI), Universitas Indonesia, Jakarta, Indonesia https://orcid.org/0000-0002-4341-046X
  • Muchtaruddin Mansyur Department of Community Medicine, Faculty of Medicine, Universitas Indonesia, Jakarta, Indonesia https://orcid.org/0000-0002-3100-3269
  • Ina Susianti Timan Human Genetic Research Center, Indonesian Medical Education and Research Institute (IMERI), Universitas Indonesia, Jakarta, Indonesia; Department of Clinical Pathology, Faculty of Medicine, Universitas Indonesia, Jakarta, Indonesia
  • Yulia Ariani Human Genetic Research Center, Indonesian Medical Education and Research Institute (IMERI), Universitas Indonesia, Jakarta, Indonesia; Department of Biology, Faculty of Medicine, Universitas Indonesia, Jakarta, Indonesia
  • Ernawati Regional Health Laboratory DKI Jakarta, Indonesia
  • I Gusti Lanang Sidhiarta Department of Pediatrics, Faculty of Medicine, Universitas Udayana, Bali, Indonesia
  • Nadhifa Tazkia Ramadhani Human Genetic Research Center, Indonesian Medical Education and Research Institute (IMERI), Universitas Indonesia, Jakarta, Indonesia https://orcid.org/0009-0006-5348-4119
  • Nurul Muhammad Prakoso Human Genetic Research Center, Indonesian Medical Education and Research Institute (IMERI), Universitas Indonesia, Jakarta, Indonesia https://orcid.org/0000-0003-1864-8676
  • Damayanti Rusli Sjarif Department of Pediatrics, Faculty of Medicine, Universitas Indonesia, Cipto Mangunkusumo Hospital, Jakarta, Indonesia; Human Genetic Research Center, Indonesian Medical Education and Research Institute (IMERI), Universitas Indonesia, Jakarta, Indonesia
Keywords: buccal, DNA, isolation, quality, swabs
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Abstract

BACKGROUND Genomic medicine has great potential for diagnoses, disease prediction, and targeted treatment. Buccal swabs are a suitable non-invasive method for neonates to obtain DNA samples. Due to Indonesia's geographical conditions, samples require a prolonged time to reach the genetic laboratory. This study aimed to compare the DNA quality of buccal swabs in neonates between immediate and after-storage extraction.

METHODS This study was part of a study about the profile of human milk oligosaccharide and FUT2 genotype in Indonesian mother-infant dyads consisting of 20 neonates. 1 swab stick for each participant was taken using a standardized buccal swabbing protocol and divided into 2 isovolume aliquots, which were grouped into the immediate (extraction was performed within 3 days after sampling) and storage groups (extraction was performed on the 14th day after storage in 4°C). DNA yield and purity A260/280 ratio were measured by spectrophotometry. The PCR amplification and Sanger sequencing were performed to validate the DNA isolate quality for downstream application.

RESULTS The DNA yield for the immediate group was similar compared with the storage group (9.50 [4.89] versus 9.10 [5.05] µg), p = 0.659, as well as DNA purity A260/280 (1.58 [0.24] versus 1.56 [0.28]), p = 0.785. PCR and sequencing of FUT2 results also showed similar quality between both groups.

CONCLUSIONS The similar DNA quality and sequencing results between immediate and storage extraction confirmed that buccal swabs could be stored for 2 weeks, allowing ample time for sample shipping from remote areas to the laboratory.

References

  1. Van-Wieren-de-Wijer DB, Maitland-van-der-Zee AH, de Boer A, Belitser SV, Kroon AA, de Leeuw PW, et al. Determinants of DNA yield and purity collected with buccal cell samples. Eur J Epidemiol. 2009;24(11):677-82. https://doi.org/10.1007/s10654-009-9388-x

  2. Siswanto JE, Berlian T, Putricahya E, Panggalo LV, Yuniani L. [DNA isolation in blood and buccal swab samples in retinopathy of prematurity patients: comparison of the concentration and purity index]. Sari Pediatri. 2016;18(4):270-7. Indonesian. https://doi.org/10.14238/sp18.4.2016.270-7

  3. Said M, Cappiello C, Devaney JM, Podini D, Beres AL, Vukmanovic S, et al. Genomics in premature infants: a non-invasive strategy to obtain high-quality DNA. Sci Rep. 2014;4:4286. https://doi.org/10.1038/srep04286

  4. Navarro D, Durán NS, Álvarez R. A method for preserving buccal swabs samples for gDNA integrity. Biotechniques. 2016;61(3):153. https://doi.org/10.2144/000114455

  5. Rogers NL, Cole SA, Lan HC, Crossa A, Demerath EW. New saliva DNA collection method compared to buccal cell collection techniques for epidemiological studies. Am J Hum Biol. 2007;19(3):319-26. https://doi.org/10.1002/ajhb.20586

  6. Grujičić NK, Davidović S, Takić D, Mojsin M, Stevanović M. Direct PCR amplification of the HVSI region in mitochondrial DNA from buccal cell swabs. Arch Biol Sci. 2012;64(3):851-8. https://doi.org/10.2298/ABS1203851G

  7. DNA Genotek. DNA collection kits for diagnostic: easy and reliable DNA collection with ORAcollect-DNA [Internet]. DNA Genotek [cited 2022 Dec 28]. Available from: https://www.dnagenotek.com/ROW/products/collection-human/oracollect-dna/OCR-100.html.

  8. Thermo Fisher Scientific. Microplate readers [Internet]. DNA Genotek [cited 2022 Dec 28]. Available from: https://www.thermofisher.com/id/en/home/life-science/lab-equipment/microplate-instruments/plate-readers.html.

  9. Ghatak S, Muthukumaran RB, Nachimuthu SK. A simple method of genomic DNA extraction from human samples for PCR-RFLP analysis. J Biomol Tech. 2013;24(4):224-31. https://doi.org/10.7171/jbt.13-2404-001

  10. Lefebvre G, Shevlyakova M, Charpagne A, Marquis J, Vogel M, Kirsten T, et al. Time of lactation and maternal fucosyltransferase genetic polymorphisms determine the variability in human milk oligosaccharides. Front Nutr. 2020;7:574459. https://doi.org/10.3389/fnut.2020.574459

  11. Meridian Bioscience. MyTaqTM DNA polymerases [Internet]. Meridian Bioscience [cited 2022 Dec 28]. Available from: https://www.bioline.com/mytaq.

  12. Wittmeier P, Hummel S. Agarose gel electrophoresis to assess PCR product yield: comparison with spectrophotometry, fluorometry and qPCR. Biotechniques. 2022;72(4):155-8. https://doi.org/10.2144/btn-2021-0094

  13. Mulot C, Stücker I, Clavel J, Beaune P, Loriot MA. Collection of human genomic DNA from buccal cells for genetics studies: comparison between cytobrush, mouthwash, and treated card. J Biomed Biotechnol. 2005;2005(3):291-6. https://doi.org/10.1155/JBB.2005.291

  14. Livy A, Lye S, Jagdish CK, Hanis N, Sharmila V, Ler LW, et al. Evaluation of quality of DNA extracted from buccal swabs for microarray based genotyping. Indian J Clin Biochem. 2012;27(1):28-33. https://doi.org/10.1007/s12291-011-0154-y

  15. Prakoso N, Priambodo R, Ariani, Y, Hafifah C, Sjarif D. Identification of a novel variant in exon 5 of galactosamine (N-acetyl)-6-sulfatase gene in mucopolysaccharidosis IVA patients in Indonesia. J Nat Sci Biol Med. 2019;10(3):S99-102.

  16. Dilhari A, Sampath A, Gunasekara C, Fernando N, Weerasekara D, Sissons C, et al. Evaluation of the impact of six different DNA extraction methods for the representation of the microbial community associated with human chronic wound infections using a gel-based DNA profiling method. AMB Express. 2017;7(1):179. https://doi.org/10.1186/s13568-017-0477-z

  17. Ghaheri M, Kahrizi D, Yari K, Babaie A, Suthar RS, Kazemi E. A comparative evaluation of four DNA extraction protocols from whole blood sample. Cell Mol Biol (Noisy-le-grand). 2016;62(3):120-4.

  18. National Library of Medicine. dbSNP short genetic variations: reference SNP (rs) report rs281377 [Internet]. Bethesda: National Center for Biotechnology Information; 2022 [cited 2023 Mar 9]. Available from: https://www.ncbi.nlm.nih.gov/snp/rs281377.

  19. National Library of Medicine. dbSNP short genetic variations: reference SNP (rs) report rs1047781 [Internet]. National Center for Biotechnology Information; 2022 [cited 2023 Mar 9]. Available from: https://www.ncbi.nlm.nih.gov/snp/rs1047781.

Published
2023-07-11
How to Cite
1.
Yuliarti K, Mansyur M, Timan IS, Ariani Y, Ernawati, Sidhiarta IGL, Ramadhani NT, Prakoso NM, Sjarif DR. DNA quality from buccal swabs in neonates: comparison of different storage time. Med J Indones [Internet]. 2023Jul.11 [cited 2024May6];32(1):7-12. Available from: http://mji.ui.ac.id/journal/index.php/mji/article/view/6899
Section
Basic Medical Research