Error rate for HLA-B antigen assignment by serology: implications for proficiency testing and utilization of DNA-based typing methods.
Until recently, the majority of HLA class I typing has been performed by serology. Expensive commercial typing trays are frequently used for testing non-Caucasian subjects and new strategies using DNA-based methods have been adopted for improving clinical histocompatibility testing results and adapted as supplements in proficiency testing. A double-blind comparison of the typing of HLA-B specificities in 40 samples was carried out between serology and two polymerase chain reaction (PCR) methods, PCR amplification with sequence-specific primers (PCR-SSP) and PCR amplification and subsequent hybridization with sequence-specific oligonucleotide probes (PCR-SSOP). The results demonstrated 22.5% misassignments of HLA-B antigens by serology. There was complete concordance between the results obtained with the two PCR based typing methods. A second panel of 20 donor samples with incomplete or ambiguous serologic results was analyzed by PCR-SSP and SSOP Both PCR methods identified correctly the HLA-B antigens. Our results suggest that more accurate typing results can be achieved by complementing serologic testing with DNA-based typing techniques. The level of resolution for HLA-B antigen assignment can be obtained by this combination of serology and limited DNA-based typing is equivalent to the HLA-B specificities defined by the WHO-HLA Committee. This level of resolution cannot routinely be achieved in clinical histocompatibility testing or in proficiency testing using serologic reagents only.
Bozón
M V
MV
Dana-Farber Cancer Institute, Department of Pathology, Harvard Medical School, Boston, Massachusetts 02115, USA.
Delgado
J C
JC
Selvakumar
A
A
Clavijo
O P
OP
Salazar
M
M
Ohashi
M
M
Alosco
S M
SM
Russell
J
J
Yu
N
N
Dupont
B
B
Yunis
E J
EJ
D004247:DNA / Q000235:genetics*; D017931:DNA Primers / Q000235:genetics; D015343:DNA Probes, HLA / Q000235:genetics*; D003951:Diagnostic Errors; D005069:Evaluation Studies as Topic; D005805:Genes, MHC Class I; D005838:Genotype; D015235:HLA-B Antigens / Q000032:analysis* / Q000235:genetics; D006650:Histocompatibility Testing / Q000379:methods* / Q000706:statistics & numerical data; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D044469:Racial Groups / Q000235:genetics; D011933:Reagent Kits, Diagnostic; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D012698:Serologic Tests / Q000706:statistics & numerical data
D017931:DNA Primers; D015343:DNA Probes, HLA; D015235:HLA-B Antigens; D011933:Reagent Kits, Diagnostic; D004247:DNA