Abstract
BackgroundUrine 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.MethodsIn 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.ResultsHLA 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.ConclusionsThis 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.
Highlights
Two-thirds of all kidney transplant recipients in Hong Kong underwent transplantation overseas [1]
We previously reported a novel method to determine mismatched donor HLA using fresh or formalin-fixed paraffin-embedded renal allograft tissue [3,4]
We demonstrated that the allograft tissue expressed both donor and recipient HLA, and mismatched donor antigens could be deduced by subtracting the recipient’s HLA from the allograft HLA
Summary
Two-thirds of all kidney transplant recipients in Hong Kong underwent transplantation overseas [1]. The lack of donor information hinders the investigation for donorspecific antibody (DSA) and hampers the diagnosis and management of antibody-mediated rejection. Donors sharing HLA antigens with previously failed transplants, even in the absence of DSA in the potential recipient, are avoided in some centres [2]. To tackle this problem, we previously reported a novel method to determine mismatched donor HLA using fresh or formalin-fixed paraffin-embedded renal allograft tissue [3,4]. The information facilitates molecular diagnosis and patient management of antibody mediate rejection, and enables appropriate selection of donor kidney for retransplantation. An optimized protocol would be required to determine mis-matched donor HLA specificities in view of the scarcity of DNA obtained in some cases
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