PDF(Pubmed)
Abstract：
Previously, we demonstrated unique insertion/deletion polymorphisms of equine histidine-rich glycoprotein (eHRG) with five genotypes composed of 45-bp or 90-bp deletions in the histidine-rich region of eHRG in Thoroughbred horses. Although leukocytes are typically used to collect DNA for genotyping, blood sampling from animals is sometimes difficult and invasive. Moreover, the method for extracting DNA from blood leukocytes involves complicated steps and must be performed soon after blood sampling for sensitive gene analysis. In the present study, we performed eHRG genotyping using DNA, isolated from oral mucosa swabs collected by rubbing the mucosa on the underside of the upper lip of horses and 100 mg of freshly excreted feces obtained by scraping their surface. In the present study, we performed eHRG genotyping using DNA isolated from oral mucosa swabs and feces of horses (18 Thoroughbreds, 17 mixed breeds, 2 warm bloods), and compared the accuracy of this method with that of the method using DNA from leukocytes. The DNA derived from oral mucosa swabs was sufficient in quantity and quality for eHRG genotyping. However, DNA derived from fecal samples requires a more sensitive detection system because of contamination with non-horse DNA, and the test quality is low. Collection of oral mucosa swabs is less invasive than blood sampling; further, oral swabs can be stored for a longer period in a specified high-quality solution. Therefore, collecting DNA samples from oral mucosa swabs is recommended for the genetic analysis of not only horses but also other animals that are not accustomed to humans.
摘要：
以前,我们证明了马富含组氨酸糖蛋白(eHRG)的独特插入/缺失多态性,在纯种马的eHRG富含组氨酸区域有5种基因型,由45bp或90bp缺失组成.尽管白细胞通常用于收集DNA进行基因分型，从动物的血液采样有时是困难和侵入性的。此外，从血液白细胞中提取DNA的方法涉及复杂的步骤，必须在血液采样后不久进行，以进行敏感的基因分析。在本研究中,我们用DNA进行了eHRG基因分型,从通过摩擦马上唇下侧的粘膜收集的口腔粘膜拭子和通过刮擦表面获得的100毫克新鲜排泄的粪便中分离出来。在本研究中,我们使用从口腔粘膜拭子和马粪便中分离的DNA进行eHRG基因分型(18种纯种，17个混合品种，2个温暖的血液），并比较了该方法与使用白细胞DNA的方法的准确性。来自口腔粘膜拭子的DNA在数量和质量上都足以进行eHRG基因分型。然而,来自粪便样本的DNA需要更灵敏的检测系统，因为非马DNA污染，测试质量低。收集口腔粘膜拭子的侵入性比采血小；此外，口腔拭子可以在指定的高质量溶液中储存更长的时间。因此,建议从口腔粘膜拭子中收集DNA样本，不仅用于马的遗传分析，还用于其他不习惯人类的动物的遗传分析。
