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J. Xu, Y. Xu, N. Liu; The Use of Buccal Cells as a Noninvasive Source of Genomic DNA for Genetic Screening in Patients With Age-Related Macular Degeneration. Invest. Ophthalmol. Vis. Sci. 2009;50(13):1608.
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© ARVO (1962-2015); The Authors (2016-present)
Blood samples are an excellent source of large amounts of genomic DNA. However, alternative sources are often needed for epidemiological studies aiming to evaluate the role of genetic factors in human disease susceptibility. Buccal swabs are the most commonly used protocols for buccal cell collection. The purpose of this study was to evaluate the protocol of buccal swabs as a noninvasive method for obtaining genomic DNA in screening risk polymorphisms in patients with exudative age-related macular degeneration (AMD).
Blood and buccal swabs were collected from 65 patients with exudative AMD. Genomic DNA was isolated from blood and buccal swabs. Yields of genomic DNA from either blood or buccal swabs were determined by photometry. With the genomic DNA obtained from blood or buccal cells, genotyping was performed for complement factor H (CFH), LOC387715, and HTRA1 polymorphisms using a method of polymerase chain reaction followed by restriction enzyme digestion.
Both blood and buccal swabs provided adequate amounts and high quality of genomic DNA for PCR assay. The quantities of DNA obtained were 3.94±1.04 µg from buccal swab of the upper/inferior gingival mucosa and 3.17±1.46 µg from buccal swab of the left or right side of the buccal mucosa. The quantity of DNA from buccal swab of the buccal mucosa stored at -20°C for 12months was 3.10±1.17 µg. In all 65 patients, genomic DNA isolated from buccal swabs revealed exactly the same results as that originated from blood regarding the CFH, LOC387715, and HTRA1 Polymorphisms.
Our data suggest that the buccal swabs are an adequate, noninvasive, and reliable source of genomic DNA for genetic screening. The yields of genomic DNA from buccal swabs of the upper/inferior gingival mucosa were higher than those of the left or right side of the buccal mucosa. Swabs storedfor 12 months at -20°C are still stable sources of genomic DNA.
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