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Arthritis
Volume 2013 (2013), Article ID 504109, 4 pages
http://dx.doi.org/10.1155/2013/504109
Research Article

Application of a Simple In-House PCR-SSP Technique for HLA-B* 27 Typing in Spondyloarthritis Patients

1National Institute of Nutrition, ICMR, Jamai Osmania, Tarnaka, Hyderabad, Andhra Pradesh, India
2Department of Medicine, King Edward Memorial Hospital Campus, Parel, Mumbai 400012, India
3National Institute of Immunohematology, 13th Floor, NMS Building, King Edward Memorial Hospital Campus, Parel, Mumbai, Maharashtra 400012, India

Received 3 October 2013; Accepted 12 November 2013

Academic Editor: Ruben Burgos-Vargas

Copyright © 2013 Devraj J. Parasannanavar et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

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.