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. 2024 Dec 21;16(12):e76140.
doi: 10.7759/cureus.76140. eCollection 2024 Dec.

A Modified Method for DNA Extraction From Buccal Cells for Genetic Epidemiological Studies Among Children

Wendy Yesenia Escobar de González  1 , Katleen Argentina Aguirre de Rodríguez  1 , Vianney Castañeda Monroy  2 , Nuria Patiño-Marín  3 , Carlo E Medina-Solís  4   5 , Nereyda Niño-Martínez  3 , Yolanda Terán-Figueroa  3
Affiliations

A Modified Method for DNA Extraction From Buccal Cells for Genetic Epidemiological Studies Among Children

Wendy Yesenia Escobar de González et al. Cureus. .

Abstract

Background: In large-scale molecular studies, a protocol that generates high yields and quality DNA for future polymerase chain reaction (PCR) assays is needed. The collection of buccal cells by cytobrush may represent an efficient, noninvasive, and inexpensive method for obtaining genetic material from school populations. The aim of this study was to develop a method to obtain genomic DNA from buccal cells of schoolchildren, and the DNA was extracted immediately after collecting the buccal cell samples and after storing the samples for 8 months at -20 °C to establish the feasibility of the method for epidemiological studies.

Methods: Forty-five Salvadoran schoolchildren aged between seven and fifteen years participated. Two samples of buccal cells were collected with cytobrush from each subject. The yield of the extracted DNA was evaluated by spectrophotometry, and purity was measured using optical density (OD) 260/280. The functional quality was assessed by PCR and established by amplifying the 536 bp region of the β-hCG gene.

Results: The yield was 65.70 μg for the immediate extraction group and 47.63 μg for the extraction group after eight months of storage at -20°C. The purity measured was 2.0 for immediate extraction and 1.9 for extraction after eight months. The functional quality was greater than 90% for both groups.

Conclusion: This protocol can be used to obtain DNA of high yield, purity, and functional quality from schoolchildren for genetic epidemiological studies based on PCR. This method is effective whether DNA is extracted immediately after collection or after buccal cell samples have been stored at 20°C for eight months.

Keywords: dna; epidemiology; genetics; molecular biology; pcr test.

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Conflict of interest statement

Human subjects: Consent for treatment and open access publication was obtained or waived by all participants in this study. Ethics Committee of the Faculty of Dentistry of the University of El Salvador issued approval CEI-FOUES-2022-012. We declare that this research has been approved by the Ethics Committee of the Faculty of Dentistry of the University of El Salvador, according to the resolution CEI-FOUES-2022-012. Animal subjects: All authors have confirmed that this study did not involve animal subjects or tissue. Conflicts of interest: In compliance with the ICMJE uniform disclosure form, all authors declare the following: Payment/services info: All authors have declared that no financial support was received from any organization for the submitted work. Financial relationships: All authors have declared that they have no financial relationships at present or within the previous three years with any organizations that might have an interest in the submitted work. Other relationships: All authors have declared that there are no other relationships or activities that could appear to have influenced the submitted work.

Figures

Figure 1
Figure 1. 1% electrophoresis gel of genomic DNA from buccal cell DNA obtained by cytobrush.
Lane 1 and 20: DNA marker of 100 bp. Immediate extraction: 1A, 2A, 3A, 4A, 5A, 6A, 7A, 8A and 9A. Extraction after eight months of storage at -20°C: 1B, 2B, 3B, 4B, 5B, 6B, 7B, 8B, and 9B.
Figure 2
Figure 2. Verification of human β-globin gene PCR products on a 2% agarose gel. Lane 1 and 12: DNA marker of 100 bp.
Immediate extraction: 1A, 2A, 3A, 4A and 5A. Extraction after eight months of storage at -20°C: 1B, 2B, 3B, 4B and 5B.
See this image and copyright information in PMC

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