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首页> 外文期刊>Journal of Immunological Methods >Saliva: a convenient source of DNA for analysis of bi-allelic polymorphisms of Fc gamma receptor IIA (CD32) and Fc gamma receptor IIIB (CD16).
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Saliva: a convenient source of DNA for analysis of bi-allelic polymorphisms of Fc gamma receptor IIA (CD32) and Fc gamma receptor IIIB (CD16).

机译：唾液：方便的DNA来源，用于分析Fcγ受体IIA（CD32）和Fcγ受体IIIB（CD16）的双等位基因多态性。

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摘要

Genetic polymorphisms of low-affinity IgG Fc receptors (Fc gamma R) have been found to influence binding of human IgG subclass antibodies, and may influence susceptibility to certain types of infectious and autoimmune diseases. Phenotypic and/or genotypic analyses of Fc gamma R polymorphisms have traditionally employed peripheral venous blood as a source of leukocytes or genomic DNA, respectively. The present study was undertaken to determine whether human salivary DNA is a suitable alternative to DNA extracted from blood for genetic analysis of FC gamma R allelic polymorphisms. Genomic DNA was extracted from whole saliva of 69 healthy adult volunteers using a commercial DNA purification kit. The average quantity of genomic DNA isolated per ml of saliva was 19.2 +/- 14.1 micrograms. To assess intrasubject variation in yield of salivary DNA, ten saliva samples were collected from a single donor over a 3-month period. The average yield of DNA recovered from these samples was 25.2 +/- 13.7 micrograms. Volumes of saliva as small as 100 microliters, as well as saliva samples stored at -70 degrees C for prolonged periods (up to 6 years), provided DNA in amounts sufficient for PCR-based genetic analysis. Two comparative PCR assays were performed using DNA extracted from both peripheral blood and saliva from a number of individuals. The assays were able to detect a single nucleotide substitution (G-->A) in the Fc gamma RIIA gene, as well as two codominant alleles encoding the NA polymorphism in Fc gamma RIIIB, respectively. Furthermore, Fc gamma RIIA and Fc gamma RIIIB genotype results were confirmed by quantitative flow cytometry using specific monoclonal antibodies. Complete concordance was achieved between the typing results of our salivary DNA, and blood DNA-based assays. Therefore, saliva appears to be an excellent source of DNA for studies of Fc gamma RIIA and Fc gamma RIIIB polymorphisms.
机译：已发现低亲和力IgG Fc受体（Fc gamma R）的遗传多态性会影响人IgG亚类抗体的结合，并可能影响对某些类型的传染性和自身免疫性疾病的易感性。 FcγR多态性的表型和/或基因型分析传统上分别采用外周静脉血作为白细胞或基因组DNA的来源。进行本研究以确定人唾液DNA是否适合替代从血液中提取的DNA，用于FCγR等位基因多态性的遗传分析。使用商业DNA纯化试剂盒从69名健康成人志愿者的整个唾液中提取基因组DNA。每毫升唾液中分离出的基因组DNA的平均量为19.2 +/- 14.1微克。为了评估受试者体内唾液DNA产量的变化，在3个月的时间内从一位供体中收集了十份唾液样品。从这些样品中回收的DNA的平均产量为25.2 +/- 13.7微克。唾液体积小至100微升，以及在-70摄氏度下长时间（长达6年）存储的唾液样品，提供了足以用于基于PCR的遗传分析的DNA量。使用从许多个体的外周血和唾液中提取的DNA进行了两次比较PCR分析。该测定法能够检测FcγRIIA基因中的单个核苷酸取代（G→A）以及两个分别编码FcγRIIIB中NA多态性的共性等位基因。此外，通过使用特异性单克隆抗体的定量流式细胞术证实了FcγRIIA和FcγRIIIB基因型结果。唾液DNA的分型结果与基于血液DNA的检测结果完全一致。因此，唾液似乎是研究FcγRIIA和FcγRIIIB多态性的优良DNA来源。

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