%0 Journal Article
%T Application of a Simple In-House PCR-SSP Technique for HLA-B* 27 Typing in Spondyloarthritis Patients
%A Devraj J. Parasannanavar
%A Anjali Rajadhyaksha
%A Kanjaksha Ghosh
%J Arthritis
%D 2013
%I Hindawi Publishing Corporation
%R 10.1155/2013/504109
%X Background. Microlymphocytotoxicity (MLCT) and flowcytometry (FC) are the conventional serological methods to detect HLA-B* 27. Due to some disadvantages in these methods, most of the HLA laboratories have now switched over to molecular methods. Molecular techniques based on commercial kits are expensive; as such many laboratories with limited funds in developing countries cannot afford these techniques. Aims. Our main aim was to standardize a simple inexpensive in-house PCR-SSP technique for HLA-B* 27 typing. Materials and Methods. Sequence Specific primers were designed to amplify all the subtypes of B* 27 using IMGT-HLA sequence database. Accuracy was checked by retyping of 90 PCR-SSOP typed controls. Results. The presence of 149£¿bp specific band with control band on 2% agarose gel showed B* 27 positivity. No discrepancies were found when compared with PCR-SSOP results. The frequency of HLA-B* 27 was found to be significantly increased (68.75% versus 4.40%, O.R 46.909: value ) among 700 SpA patients as compared to controls. Clinically, 54% of patients had polyarticular arthritis with SI joints involvement (68%) and restricted spine flexion (60%). Conclusion. In-house PCR-SSP technique is very simple and inexpensive technique to detect B* 27 allele, which was strongly associated with SpA patients from Western India. 1. Introduction Human leucocyte antigen-B27 is a major histocompatibility complex class I molecule that is strongly associated with AS and related seronegative spondyloarthritis (SpA). Ankylosing spondylitis (AS) is associated with B27 with a relative risk of 95 which is the highest among all HLA disease associations [1]. The association of HLA-B27 with AS was first reported in 1973 [2, 3] and confirmed with related SpA later by many other investigators [4¨C7]. The frequency of HLA-B27 ranges from 80% to 90% among patients as compared to 4¨C8% in controls. This high association stimulated considerable interest in HLA-B* 27 testing as a diagnostic tool in these inflammatory diseases. The frequency of HLA-B27 in AS or other related SpA among Indian population varies from 40% to 80% as compared to 1.4¨C8% of controls as shown in Table 1 [8¨C15]. Techniques generally employed for the routine typing of HLA-B27 are the microlymphocytotoxicity test (MLCT) [16] and Flow cytometry (FC) [17]. Both these tests rely on the detection of cell surface antigens by antibodies. There are some disadvantages in MLCT like requirement of viable cells, cross-reactive nature of HLA antigens, unavailability of B* 27 specific antisera which cover all HLA-B* 27 alleles
%U http://www.hindawi.com/journals/arthritis/2013/504109/