Abstract:

Purpose: In the present study, we aimed to develop a reliable, cost effective and suitable for routine use in-house HLA-B*27 PCR-SSP typing kit for the molecular diagnosis of Ankylosing spondylitis and other spondyloarthropathies.  Material and Methods: The study included 565 cases with 446 negative and 119 positive HLA-B*27 subjects. PCR optimization was performed for HLA-B*27 alleles using sequence-specific and internal control primers designed based on HLA-B and beta globin gene sequences, respectively.  Results: The presence of 149 bp specific band with 273 bp internal control band showed B*27 positivity and the presence of only 273 bp control band showed B*27 negativity on 2.0% agarose gel. The HLA-B*27 results show 100% sensitivity and specificity to our in-house SSP kit developed by our laboratory and the commercially available SSP kits. The cost of the in-house PCR-SSP kit we developed was at least 50 times cheaper than the commercial kits. In addition to the faultless correlation between the kits, our in-house SSP kit did not cross-react with other HLA-B alleles such as B*07, B*12, B*13, B*16, B*17, B*22, B*37, B*40, B*41, B*42, B*47 and B*48. Conclusion: The fully evaluated findings show that in-house SSP kit for HLA-B*27 typing we developed is reliable, cost effective and suitable for routine use in tissue typing and genetic laboratories.

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