A Modified Protocol with Improved Detection Rate for Mis-Matched Donor HLA from Low Quantities of DNA in Urine Samples from Kidney Graft Recipients

• Published in: PloS one Vol. 11; no. 11; p. e0166427
• Main Authors: Kwok, Janette, Choi, Leo C W, Ho, Jenny C Y, Chan, Gavin S W, Mok, Maggie M Y, Lam, Man-Fei, Chak, Wai-Leung, Cheuk, Au, Chau, Ka-Foon, Tong, Matthew, Chan, Kwok-Wah, Chan, Tak-Mao
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 18.11.2016; Public Library of Science (PLoS)
• Subjects: Alleles; Antigens; Biology and Life Sciences; Deduction; Deoxyribonucleic acid; DNA; DNA - urine; Drb1 protein; Genetic research; Genetic testing; Geriatrics; Graft Rejection - genetics; Graft Rejection - immunology; Graft Survival - genetics; Graft Survival - immunology; Histocompatibility antigen HLA; Histocompatibility Testing - methods; HLA antigens; HLA Antigens - genetics; HLA Antigens - immunology; Humans; Kidney transplantation; Kidney Transplantation - adverse effects; Kidneys; Laboratories; Medical research; Medicine; Medicine and Health Sciences; Organ transplant recipients; Pathology; Patients; Phenotypes; Polymerase Chain Reaction; Primers; Research and analysis methods; Time Factors; Tissue Donors; Tissue typing; Transplant Recipients; Transplants & implants; Urine; Hong Kong China; China
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0166427

Urine from kidney transplant recipient has proven to be a viable source for donor DNA. However, an optimized protocol would be required to determine mis-matched donor HLA specificities in view of the scarcity of DNA obtained in some cases. In this study, fresh early morning urine specimens were obtained from 155 kidney transplant recipients with known donor HLA phenotype. DNA was extracted and typing of HLA-A, B and DRB1 loci by polymerase chain reaction-specific sequence primers was performed using tailor-made condition according to the concentration of extracted DNA. HLA typing of DNA extracted from urine revealed both recipient and donor HLA phenotypes, allowing the deduction of the unknown donor HLA and hence the degree of HLA mis-match. By adopting the modified procedures, mis-matched donor HLA phenotypes were successfully deduced in all of 35 tested urine samples at DNA quantities spanning the range of 620-24,000 ng. This urine-based method offers a promising and reliable non-invasive means for the identification of mis-matched donor HLA antigens in kidney transplant recipients with unknown donor HLA phenotype or otherwise inadequate donor information.