Human leucocyte antigen typing: techniques and technology, a critical appraisal

Area of Science:

• Immunogenetics
• Molecular Biology
• Genomics

Background:

• Human Leukocyte Antigen (HLA) identification has evolved significantly since the 1960s.
• Early methods relied on serological reagents, with Polymerase Chain Reaction (PCR) revolutionizing HLA genotyping.

Purpose of the Study:

• To review the evolution of HLA typing methodologies.
• To discuss the merits and disadvantages of current HLA typing technologies.
• To explore future trends and challenges in HLA typing.

Main Methods:

• Review of historical and current HLA typing techniques.
• Analysis of limitations of traditional methods like PCR-Restriction Fragment Length Polymorphism (RFLP) and Reference Strand Mediated Conformation Analysis (RSCA).
• Evaluation of contemporary methods such as Sequence-Specific Primer PCR and Sequencing Based Typing (SBT) in light of Next Generation Sequencing (NGS).

Main Results:

• Older techniques (PCR-RFLP, RSCA) are obsolete due to HLA polymorphism.
• Current routine methods (Sequence-Specific Primer PCR, SBT) are increasingly challenged by high-resolution HLA variation.
• Next Generation Sequencing (NGS) represents the leading edge, with future technologies poised to further advance HLA typing.

Conclusions:

• The field of HLA typing is characterized by rapid technological advancement.
• Increasing HLA polymorphism necessitates continuous evolution of typing methodologies.
• Future HLA typing will likely be dominated by high-throughput sequencing technologies and innovative approaches to manage complex genetic variation.