The aim of the present study was to value the possible application of the ARMS method in clinical practice ofkidney transplantation and to compare this allelic resolution with serologic techniques. For this purpose, we analyzedthe incidence of HLA-A and -B serologic blanks in cadaveric donors, and we used the PCR-ARMS techniqueto discriminate whether they reflected a homozygous formor a mistyped allelic assignation. PCR-ARMS seems to be a method with an useful applicability in definition of class I compatibility in cadaverickidney transplant for its speed of execution (4 hours including DNA extraction), ease of result interpretation,and the best resolution in allele definition, even if for the present it is limited by relatively low resolution capabilityand by the difficulty in distinguishing some alleles and some heterozygote forms, depending on the identical reactivitiesof some primer mixes for certain allele combinations. It would be advisable to raise the number of mixes available toavoid these problems and to have a higher resolution, expecially for HLA-B antigens.The better definition of HLA-A locus obtained by this molecular method to serologic typing concerned the A19group, cases in which one allele might mask the presence of the second cross-reactive allele, such as Al 1 missed allele ina donor serologically typed as Al/blank or A24 missed allele in a A2/blank donor or A23,24 in a A9/blank case.Moreover, antigens that one would expect to identify easily by serology, such as Al or A2, can be missed but detectedby DNA typing, probably for a low expression of a particular antigen.As regards HLA-B, a more polymorphic locus thanHLA-A, it is very important to obtain a better definitionbecause these antigens are mainly involved, together with HLA class II antigens, in the clinical outcome of thetransplant. Therefore, the increase of HLA-B mismatches, found retrospectively in our donor-recipient pairs, underlinesthe importance of PCR-ARMS and suggests its possible application in routine cadaver donor typing.
Comparison between HLA class I PCR-ARMS and serologic typing in cadaveric kidney transplantation.
Adorno D;Canossi A;Ozzella G;Di Rocco M;Liberatore G;
1997
Abstract
The aim of the present study was to value the possible application of the ARMS method in clinical practice ofkidney transplantation and to compare this allelic resolution with serologic techniques. For this purpose, we analyzedthe incidence of HLA-A and -B serologic blanks in cadaveric donors, and we used the PCR-ARMS techniqueto discriminate whether they reflected a homozygous formor a mistyped allelic assignation. PCR-ARMS seems to be a method with an useful applicability in definition of class I compatibility in cadaverickidney transplant for its speed of execution (4 hours including DNA extraction), ease of result interpretation,and the best resolution in allele definition, even if for the present it is limited by relatively low resolution capabilityand by the difficulty in distinguishing some alleles and some heterozygote forms, depending on the identical reactivitiesof some primer mixes for certain allele combinations. It would be advisable to raise the number of mixes available toavoid these problems and to have a higher resolution, expecially for HLA-B antigens.The better definition of HLA-A locus obtained by this molecular method to serologic typing concerned the A19group, cases in which one allele might mask the presence of the second cross-reactive allele, such as Al 1 missed allele ina donor serologically typed as Al/blank or A24 missed allele in a A2/blank donor or A23,24 in a A9/blank case.Moreover, antigens that one would expect to identify easily by serology, such as Al or A2, can be missed but detectedby DNA typing, probably for a low expression of a particular antigen.As regards HLA-B, a more polymorphic locus thanHLA-A, it is very important to obtain a better definitionbecause these antigens are mainly involved, together with HLA class II antigens, in the clinical outcome of thetransplant. Therefore, the increase of HLA-B mismatches, found retrospectively in our donor-recipient pairs, underlinesthe importance of PCR-ARMS and suggests its possible application in routine cadaver donor typing.File | Dimensione | Formato | |
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Transpl. Proc 1997 HLA-I.pdf
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Descrizione: Comparison between HLA class I PCR-ARMS and serologic typing in cadaveric kidney transplantation
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