[使用口腔拭子作为年龄相关性黄斑变性患者基因筛查的基因组DNA来源]

Xu Yu-le, Zhou Hai-ying, Xu Jun, Liu Ning-pu

Capital Medical University, Beijing, China.

Zhonghua Yan Ke Za Zhi. 2012 Feb;48(2):114-8.

OBJECTIVE

The collection of buccal cells with swabs provides a noninvasive method for obtaining genomic DNA for genetic screening. We aimed to study the feasibility of using buccal swabs for genetic screening in patients with exudative age-related macular degeneration (AMD).

METHODS

Blood and buccal swabs were collected for genetic analysis from 65 patients with exudative AMD. Genomic DNA was isolated from either blood or buccal swabs. The yield of genomic DNA from both sources was determined by spectrophotometer. Genotyping for CFH, LOC387715, and HTRA1 Polymorphisms was performed using a method of polymerase chain reaction (PCR) followed by restriction enzyme digestion. Results using genomic DNA from blood or buccal swabs were compared.

RESULTS

Three swabs were obtained from each patient, 2 from each side of buccal mucosa, and 1 from both upper and inferior gingival mucosa. From swabs with genomic DNA extracted within a week after sample collection, an average of (3.17 ± 1.46) µg genomic DNA was obtained from swab collected from the left or right side buccal mucosa, and (3.94 ± 1.04) µg from swab collected from the upper and inferior gingival mucosa, with relatively higher yield of genomic DNA from the upper and inferior gingival mucosa (t = 6.79, P < 0.05). From swabs of the left or right side buccal mucosa after being stored at -20°C for 12 months, an average of (3.10 ± 1.17) µg genomic DNA was obtained, which showed no statistically significant difference as compared to the yield of genomic DNA extracted from newly collected swabs (t = 0.59, P > 0.05). In all 65 patients, genomic DNA isolated from either buccal swabs or blood samples showed exactly the same results regarding the genotypes of CFH, LOC387715, and HTRA1 Polymorphisms.

CONCLUSIONS

Buccal swab is a simple, noninvasive, and reliable source for obtaining genomic DNA. Swabs stored for 12 months at -20°C provide similar amount of genomic DNA as the freshly collected swabs.

目的

用拭子采集颊细胞为基因筛查提供了一种获取基因组DNA的非侵入性方法。我们旨在研究使用颊拭子对渗出性年龄相关性黄斑变性（AMD）患者进行基因筛查的可行性。

方法

从65例渗出性AMD患者中采集血液和颊拭子用于基因分析。从血液或颊拭子中分离基因组DNA。用分光光度计测定两种来源的基因组DNA产量。使用聚合酶链反应（PCR）方法随后进行限制性酶切来进行CFH、LOC387715和HTRA1多态性的基因分型。比较使用血液或颊拭子中的基因组DNA得到的结果。

结果

每位患者采集3个拭子，颊黏膜两侧各2个，上下牙龈黏膜各1个。在样本采集后一周内提取基因组DNA的拭子中，从左侧或右侧颊黏膜采集的拭子平均获得（3.17±1.46）μg基因组DNA，从上下牙龈黏膜采集的拭子平均获得（3.94±1.04）μg，上下牙龈黏膜的基因组DNA产量相对较高（t = 6.79，P < 0.05）。在-20°C储存12个月后，从左侧或右侧颊黏膜拭子中平均获得（3.10±1.17）μg基因组DNA，与从新采集拭子中提取的基因组DNA产量相比，差异无统计学意义（t = 0.59，P > 0.05）。在所有65例患者中，从颊拭子或血液样本中分离的基因组DNA在CFH、LOC387715和HTRA1多态性的基因型方面显示出完全相同的结果。