title: Assessment of fidelity and utility of the whole-genome amplification for the clinical tests offered in a histocompatibility and immunogenetics laboratory.
abstract: Increasing emphasis on the use of molecular tests in a histocompatibility and immunogenetics laboratory (HIL) poses a potential problem of lack of sufficient DNA to perform multiple genetic analyses. In this study, we report the feasibility, fidelity and utility of multiple displacement amplification (MDA) method to perform whole-genome amplification (WGA) to generate DNA specimens that can be analyzed by multiple molecular techniques and can be used for different clinical tests offered by an HIL. The MDA-generated DNA when compared with the native DNA showed 100% congruency in genotyping of 37 genes/loci using multiple downstream molecular techniques: sequence-based typing and sequence-specific primer-based typing for 5 human leukocyte antigen (HLA) class I and II genes (HLA-A, B, C, DRB1 and DQB1), luminex-based sequence-specific oligonucleotide (SSO) genotyping for a panel of 16 killer immunoglobulin-like receptor (KIR) genes and automated fragment size analysis for a panel of 15 short tandem repeat (STR) loci and amelogenin gene. For post-allogeneic hematopoietic cell transplantation (HCT) chimerism analysis, MDA-generated DNA appeared useful for enriching pre-transplant DNA but not for enriching post-transplant chimeric DNA. Overall, our results show that MDA-based WGA could generate DNA of high yield and fidelity that can be used for various clinical tests and research purposes.
authors.affiliation: Tissue Typing Laboratory, Calgary Laboratory Services, Calgary, Alberta, Canada. fkhan@ucalgary.ca
authors.forename: F
authors.initials: F
authors.lastname: Khan
authors.forename: A
authors.initials: A
authors.lastname: Liacini
authors.forename: E
authors.initials: E
authors.lastname: Arora
authors.forename: S
authors.initials: S
authors.lastname: Wang
authors.forename: M
authors.initials: M
authors.lastname: Assad
authors.forename: J
authors.initials: J
authors.lastname: Doulla
authors.forename: R M
authors.initials: RM
authors.lastname: Faridi
authors.forename: N
authors.initials: N
authors.lastname: Berka
mesh_terms: D053523:Amelogenin / Q000235:genetics / Q000276:immunology; D004247:DNA / Q000032:analysis* / Q000235:genetics; D016172:DNA Fingerprinting / Q000379:methods*; D017931:DNA Primers; D020869:Gene Expression Profiling; D056426:Genetic Loci; D015894:Genome, Human; D055106:Genome-Wide Association Study; D006680:HLA Antigens / Q000235:genetics / Q000276:immunology; D018380:Hematopoietic Stem Cell Transplantation; D006650:Histocompatibility Testing; D006801:Humans; D021141:Nucleic Acid Amplification Techniques / Q000379:methods* / Q000706:statistics & numerical data; D054340:Receptors, KIR / Q000235:genetics / Q000276:immunology; D017422:Sequence Analysis, DNA; D020080:Tandem Repeat Sequences; D014184:Transplantation, Homologous
chemical_list: D053523:Amelogenin; D017931:DNA Primers; D006680:HLA Antigens; D054340:Receptors, KIR; D004247:DNA