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Clin Biochem. 2012 Nov;45(16-17):1471-8. doi: 10.1016/j.clinbiochem.2012.05.023. Epub 2012 May 31.

Effectiveness and limitations of resolving HLA class I and class II by heterozygous ambiguity resolving primers (HARPs)--a modified technique of sequence-based typing (SBT).

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1
Department of Pathology, National Defense Medical Center, Division of Clinical Pathology, Tri-Service General Hospital, Taipei, Taiwan, ROC.

Abstract

OBJECTIVES:

The aim was to evaluate the use of combination of SBT (sequence based typing) and HARP (heterozygous ambiguity resolving primer) in HLA typing to acquire high resolution typing results.

DESIGN AND METHODS:

167 DNA samples were analyzed by SBT. The web site HARPs Finder provided by Conexio Genomics, the developer of HARPs (http://www.harpsfinder.conexio-genomics.com/index.html) was then used to search for appropriate HARPs.

RESULTS:

HARPs can resolve 95% of ambiguities for locus A; 86% for B and 60% for DRB1 locus. However, there are still limitations. Practically PCR products of un-separated alleles are used as templates for sequencing by HARP; sometimes, it is still impossible to get unambiguous typing.

CONCLUSIONS:

We outlined the advantages and disadvantages of SBT/HARP. A list of HARPs for choice to resolve ambiguity of SBT in Taiwanese population is concluded.

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