title: A Modified Protocol with Improved Detection Rate for Mis-Matched Donor HLA from Low Quantities of DNA in Urine Samples from Kidney Graft Recipients.
abstract: BACKGROUND
  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.
  
  
  METHODS
  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.
  
  
  RESULTS
  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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Division of Transplantation and Immunogenetics, Department of Pathology,Queen Mary Hospital, Hong Kong SAR, China.
authors.forename: Janette
authors.initials: J
authors.lastname: Kwok
authors.affiliation: Division of Transplantation and Immunogenetics, Department of Pathology,Queen Mary Hospital, Hong Kong SAR, China.
authors.forename: Leo C W
authors.initials: LC
authors.lastname: Choi
authors.affiliation: Division of Transplantation and Immunogenetics, Department of Pathology,Queen Mary Hospital, Hong Kong SAR, China.
authors.forename: Jenny C Y
authors.initials: JC
authors.lastname: Ho
authors.affiliation: Division of Anatomical Pathology, Department of Pathology, Queen Mary Hospital, Hong Kong SAR, China.
authors.forename: Gavin S W
authors.initials: GS
authors.lastname: Chan
authors.affiliation: Department of Medicine, The University of Hong Kong, Hong Kong SAR, China.
authors.forename: Maggie M Y
authors.initials: MM
authors.lastname: Mok
authors.affiliation: Department of Medicine, The University of Hong Kong, Hong Kong SAR, China.
authors.forename: Man-Fei
authors.initials: MF
authors.lastname: Lam
authors.affiliation: Department of Medicine, Queen Elizabeth Hospital, Hong Kong SAR, China.
authors.forename: Wai-Leung
authors.initials: WL
authors.lastname: Chak
authors.affiliation: Department of Medicine & Geriatrics, Princess Margaret Hospital, Hong Kong SAR, China.
authors.forename: Au
authors.initials: A
authors.lastname: Cheuk
authors.affiliation: Department of Medicine, Queen Elizabeth Hospital, Hong Kong SAR, China.
authors.forename: Ka-Foon
authors.initials: KF
authors.lastname: Chau
authors.affiliation: Department of Medicine & Geriatrics, Princess Margaret Hospital, Hong Kong SAR, China.
authors.forename: Matthew
authors.initials: M
authors.lastname: Tong
authors.affiliation: Department of Pathology, The University of Hong Kong, Hong Kong SAR, China.
authors.forename: Kwok-Wah
authors.initials: KW
authors.lastname: Chan
authors.affiliation: Department of Medicine, The University of Hong Kong, Hong Kong SAR, China.
authors.forename: Tak-Mao
authors.initials: TM
authors.lastname: Chan
mesh_terms: D000483:Alleles; D004247:DNA / Q000652:urine*; D006084:Graft Rejection / Q000235:genetics / Q000276:immunology; D006085:Graft Survival / Q000235:genetics / Q000276:immunology; D006680:HLA Antigens / Q000235:genetics* / Q000276:immunology; D006650:Histocompatibility Testing / Q000379:methods; D006801:Humans; D016030:Kidney Transplantation / Q000009:adverse effects; D016133:Polymerase Chain Reaction; D013997:Time Factors; D014019:Tissue Donors; D066027:Transplant Recipients
chemical_list: D006680:HLA Antigens; D004247:DNA