Saliva: a convenient source of DNA for analysis of bi-allelic polymorphisms of Fc gamma receptor IIA (CD32) and Fc gamma receptor IIIB (CD16).
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.
van Schie
R C
RC
Department of Oral Biology, School of Dental Medicine, State University of New York at Buffalo 14214-3092, USA. van_schie@sdm.buffalo.edu
Wilson
M E
ME
D000328:Adult; D000483:Alleles; D016172:DNA Fingerprinting; D005260:Female; D005820:Genetic Testing; D005838:Genotype; D006801:Humans; D008297:Male; D010641:Phenotype; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic; D017452:Receptors, IgG / Q000235:genetics*; D012463:Saliva
D017452:Receptors, IgG