Sequence-specific Primer (PCR-SSP) Technology

Analysis of the human leukocyte antigen (HLA) type is an important component of immunological research and biopharmaceutical research. Traditionally, micronucleotoxicity (MLCT) and flow cytometry (FC) were widely used to detect HLA types. Various molecular biology methods are also employed because of the limitations and deficiencies of MLCT and FC, while this expensive option is somehow difficult for cash-strapped laboratories to afford. To help scientists solve this dilemma, Creative Biolabs has standardized the sequence-specific primer (PCR-SSP) technology and applied it to HLA type detection, which can be determined by electrophoretic analysis.

Introduction to HLA Tissue Typing

HLA generally refers to the entire leukocyte antigen system, which is controlled by genes on the short arm of chromosome 6, and is part of the genetic region known as the major histocompatibility complex (MHC). The physiological role of HLA is mainly to control the self-identification of the immune system and resist the invasion of microorganisms. HLA gene ensures the accurate judgment and attack ability of the body's external threats by virtue of its extreme diversity. Because some HLA antigens are recognized on all body tissues (rather than just blood cells), the identification of HLA antigens is described as “Tissue typing”. Traditionally, HLA antigens have been classified and defined by serological techniques that rely on obtaining live lymphocyte preparations and appropriate antisera to identify the availability of HLA antigens. In the past few years, DNA technology has been widely used and gradually replaced the serological technology in clinical applications in the field of histocompatibility and immunogenetics. Many laboratories have begun to develop and apply several different DNA methods for DNA typing to detect HLA alleles.

Fig.1 Principle of PCR-SSP.

PCR-SSP for HLA Tissue Typing

The PCR-SSP technique first appeared in the early 1990s and was based on the amplification of refractory mutation systems (ARMS). The principle of this method is that a perfectly matched primer is more efficient in a PCR reaction than one or more mismatched primers. Specificity is determined by the use of sequence-specific primers, where a 3' single-base mismatch inhibits the initiation of a non-specific reaction. Due to the lack of 3' to 5' exonuclease activity of Taq polymerase, even primer pairs do not specifically anneal and are not efficiently amplified. Therefore, only the required allele will be amplified, and then the amplified product can be detected by agarose gel electrophoresis.

Creative Biolabs has developed PCR-SSP technology to provide HLA typing services to a wide range of researchers. This method costs far less than traditional analytical methods such as flow cytometry, allowing you to get experimental results in the short term and make your project more efficient.

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All listed services and products are For Research Use Only. Do Not use in any diagnostic or therapeutic applications.

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