Molecular typing for the MHC with PCR-SSP

Area of Science:

• Molecular Biology
• Immunogenetics
• Biotechnology

Background:

• Sequence-specific amplification (SSP) is a polymerase chain reaction (PCR) technique.
• It utilizes the 3'-mismatch principle for primer design to control amplification.
• Early developments occurred in the late 1980s, with significant contributions from groups at Guy's Hospital and Upjohn.

Purpose of the Study:

• To describe the principles and applications of Sequence-Specific Amplification (SSP).
• To highlight the method's utility in detecting single nucleotide polymorphisms (SNPs).
• To establish SSP as a key technique in high-resolution HLA typing for solid organ transplantation.

Main Methods:

• Designing primers with specific 3' end sequences to enable or inhibit amplification.
• Utilizing the 3'-mismatch principle to identify single nucleotide point mutations (SNPs).
• Applying PCR-SSP for high-resolution Human Leukocyte Antigen (HLA) typing.

Main Results:

• The 3'-mismatch principle allows for the detection of virtually any single nucleotide point mutation (SNP).
• SSP gained significant popularity around 1990, driven by its application in solid organ transplantation.
• SSP is now the preferred method for high-resolution HLA typing in many laboratories.

Conclusions:

• Sequence-specific amplification (SSP) is a versatile PCR-based method for SNP detection.
• Its application has expanded significantly, becoming the gold standard for high-resolution HLA typing.
• SSP has over a thousand documented applications for genes outside the Major Histocompatibility Complex (MHC).