Human leukocyte antigen (HLA)‎ typing by polymerase chain reaction-sequence specific primers (PCR-SSP)‎ in comparison with serologic methods

Abstract EN

Objective: This study was conducted to compare the efficacy and accuracy of the molecular Polymerase Chain Reaction – Sequence Specific Primers (PCR – SSP) based method with the serologic methods for HLA typing, in order to choose the best feasible test as a routine tissue typing test in the future in our clinical laboratories concerned to organs transplantation.

Methods: The HLA-I was typed serologically by lymphocytotoxicity method using (HISTO TRAY HLA-I typing Kit, Germany) and the HLA Class I by PCR-SSP method using (HISTO TYPE SSP kit, Germany).

Results: For the serologic method, the frequency of the different polymorphic HLA-I antigens were 44 for locus A, 53 for locus B and 38 for locus C in 20 screened persons (2.2, 2.6 and 1.9 polymorphic HLA antigens per single person respectively), whereas the frequency of the HLA-I (A) locus detected by PCR-SSP was 310 in 70 screened persons which makes 5.2 polymorphic (A) antigens per single person.

Also the discrepancy between the two methods was very clear in the serologic method, there was only one (A38 antigen) (2.27%) which was not detected by the PCR-SSP method, while in DNA dependent method (SSP), there were 4 (A) antigen types (A68, A31, A34, and A210).

Conclusions: Amplification of HLA loci with PCR-SSP has proved to be a rapid and accurate method for HLAA, -B and -C alleles typing.

Also PCR-SSP allowed determination of the subtypes of HLA antigens very clearly and in broader scale making the matching of HLA types easier and more precise.