Evaluation of Salivary and Oral Cell Collection Methods for Genomic DNA Extraction

Q4 Dentistry Dentistry 3000 Pub Date : 2019-07-25 DOI:10.5195/D3000.2019.91
C. A. Fernandez, F. F. C. F. Ferreira, Christiane Vasconcellos Cruz, M. C. Costa
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Abstract

The use of saliva and oral cells as sources of biological material has gained attention, due to advantages such as facility, non-invasiveness, and great patient acceptance. The objective of the study was to compare four different types of saliva and oral buccal cell collecting methods for genomic DNA extraction: (1)Expectoration of saliva, (2)Expectoration of saliva with lingual stimulation, (3)Scraping with cytological brush, and (4)Scraping with cytological brush and expectoration of saliva. The sample was composed of students and employees from the Dental School of the Federal University of Rio de Janeiro (n = 20, 10 men and 10 women with mean age of 47.60 ± 15.70 and 20.50 ± 2.1, respectively). The collections were performed with an interval of at least one day between them and the participants were instructed to stay for less than 30 minutes without eating food and brushing teeth. Samples were stored at -20°C until DNA extraction was performed using a commercially available kit (Qiagen®). Differences in DNA yield between methods were test for statistical significance with an alpha of 0.05. No sexual dimorphism was observed in relation to the concentration of DNA (p=0.76), age (p=0.91), and ethnicities (p=0.72). There was no significant difference between the collection methods in relation to the quantity and purity of the extracted DNA (p≥0.05). All methods gave lower DNA yields than the ones obtained from blood or saliva collected through comercial kits and maybe carefully use forclinical diagnostic purposes or for research experiements requiring higher DNA concentrations.
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唾液和口腔细胞采集方法用于基因组DNA提取的评价
唾液和口腔细胞作为生物材料的来源由于其便利性、非侵袭性和患者接受度高等优点而受到关注。本研究的目的是比较四种不同类型的唾液和口腔颊部细胞收集方法用于提取基因组DNA:(1)唾液的咳痰,(2)用舌刺激进行唾液的咳血,(3)用细胞学刷刮拭,以及(4)用细胞学刷子刮拭和吐唾液。样本由里约热内卢联邦大学牙科学院的学生和员工组成(n=20,10名男性和10名女性,平均年龄分别为47.60±15.70和20.50±2.1)。采集间隔至少一天,参与者被要求在不吃东西和不刷牙的情况下停留不到30分钟。样品在-20°C下储存,直到使用市售试剂盒(Qiagen®)进行DNA提取。用0.05的α检验两种方法之间DNA产量的差异是否具有统计学意义。没有观察到与DNA浓度(p=0.76)、年龄(p=0.91)和性别差异有关的两性异形,和种族(p=0.72)。在提取DNA的数量和纯度方面,两种采集方法之间没有显著差异(p≥0.05)。所有方法的DNA产量都低于通过商业试剂盒采集的血液或唾液中的DNA产量,可能会谨慎地用于临床诊断目的或需要更高DNA的研究实验浓度。
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来源期刊
Dentistry 3000
Dentistry 3000 Dentistry-Dentistry (all)
CiteScore
0.40
自引率
0.00%
发文量
25
审稿时长
19 weeks
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