Clinical efficacy of HLA B27for diagnosis of Ankylosing Spondylitis

Dr. Tahir A; Dr. Gazala T; Dr. Garg Nita

Volume : VI, Issue : IX, September - 2016

Clinical efficacy of HLA B27for diagnosis of Ankylosing Spondylitis

Dr. Tahir A, Dr. Gazala T, Dr. Garg Nita

Abstract :

Aim: The aim of our study was to find an involvement of HLA–B27 specific allele in relation to age & sex in the symptomatic & suspected cases of ankylosingspondylitis(AS) using Sequence Specific Allele PCR (SSA–PCR) method. Study design: A hospital based cross sectional study was conducted on pt’s having chronic low back ache attending the Outpatient department of Orthopaedics, Government Meical College Jammu. 67cases (47male & 20 female) in the age group (15–40 yrs) suspected of AS were selected randomly for this study. They were then divided into two groups according to age & sex 15–35 yrs & 36–60 yrs and were subjected to SSA–PCR for detection of HLA–B27. Methodology: All the 67 cases both male & females suspected of AS were selected and sequence specific allele–PCR technique was used to detect the HLA–B27 specific allele. Specimens were collected from patients attending the Out Patient Department of Orthopaedics. Whole blood samples were collected from suspected pt’s with symptoms of low backache and the samples were transported at 4°C to the laboratory for further processing. Silica column based nucleopore DNA extraction kit was utilized for high yield nucleic acid extraction. Eluted DNA was used for amplification of sequence specific allele of HLA–B27 by PCR method. Then Agarose gel electrophoresis was performed, the gel was stained with Ethediumomide which gets intercalated into DNA, and fluoresce under U.V light. After electrophoresis gel is illuminated with an U.V lamp. The ethidium omide fluresecs reddish orange in the presence of DNA, since it has intercalated with the DNA. The gel is then photographed with a digital or Polaroid camera fitted inside the gel imager. Result: The study showed that, out of 47 males included in the study 17 males (36.2%) were detected +ve with HLAB27 and 4 females (20%) out of 20 females were HLA–B27 +ve. Out of the 47 males, 24 males lying in the age group (15–35 yrs) 10 were HLA–B27 +ve & 7 out of 23 males in the age group (36–60 yrs) were +ve for HLA–B27.Of the 12 females in (15–35 yrs), 3 were +ve for HLA–B27 and 1 out of 8 females in (36–60yrs) was +ve for HLA–B27.