The Morganâ„˘ HLA SSP B27 Identification Typing Kit is for determining HLA alleles using PCR techniques with sequence specific primers (SSP). The HLA molecules of the human major histocompatibility (MHC) antigensare key elements in restricting the specificity of T-cell mediated immune responses HLA-B27 is one of the types of the HLA-B locus. The association between HLA-B27 and ankylosing spondylitis has been widely demonstrated. Ankylosing spondylitis is a form of chronic inflammation of the spine and the sacroiliac joints. The majority of patients with ankylosing spondylitis are B27positive. HLA-B27 typing provides an aid to diagnosis of ankylosing sponidylitis. Click here to see the protocol
This PCR-SSP method is based on the principle that only primers with completely matched sequences to the target sequences result in amplified products under controlled PCR conditions. The presence of amplified DNA fragment is a positive indication of the existence of allele specific sequence in the genomic DNA. On the other hand, mismatched primers do not generate amplicons. In addition to sequence specific primers, an internal control primer pair, which amplifies a conserved region of the house keeping gene, cystic fibrosis gene, is included in every PCR reaction mix, and the PCR product of the internal control primer pair serves as an indication of the integrity of PCR reaction. When the DNA fragment amplified by sequence specific primer is present, the product of the internal control primer pair is sometimes weak or absent due to the differences in amplifying efficiency between the specific primer pair and the internal control primer pair.
The PCR reaction products are examined by agarose gel electrophoresis which separates the DNA fragments by size. The amplified fragments are visualized by ethidium bromide staining followed by ultraviolet light exposure, and appear as bands. The Morganâ„˘ HLA SSP B27 Typing Kit supplies single primer mix which is designed to provide positive identification of HLA-B27 alleles.
Requires only 0.3μg DNA per test
Dried primers for easy transport
Compatible with standard 96 well PCR thermal cycler
Dual-primer-set-design: primers for internal control and B27 specific subtypes amplification
Morganâ„˘ SSPal HLA Typing Analysis Software is available for data interpretation
2 x HLA-B27 Identification Typing Trays (12 tests/tray)
3 x 1020μl Master Buffer
3 x PCR Plate Cover Membranes
1 x Gel Interpretation Worksheet
1 x Instructions for Use
*Taq is not included
*Typing Tray should be stored at 2-8oC.
*Master Buffer should be stored at -20oC.