The detection of the HLA-B27 antigen by immunomagnetic separation and enzyme-linked immunosorbent assay-comparison with a flow cytometric procedure.
The HLA-B27 antigen is an important genetic marker in ankylosing spondylitis (AS). Methods for the detection of B27 include the microlymphocytotoxicity test and, more recently, flowcytometry (FC). Here, we describe a new method, IMS-ELISA, for measuring the B27-antigen. It combines immunomagnetic separation (IMS), to obtain B27-positive cells from whole blood samples, with an enzyme-linked immunosorbent assay (ELISA) as a read-out. IMS-ELISA was tested on 367 samples obtained from five different hospitals in Taiwan. The sensitivity, specificity and accuracy of the method were compared with FC. Any conflicting data between IMS-ELISA and FC was confirmed by HLA-DNA typing via PCR-SSP (polymerase chain reaction-sequence specific primers). Overall, the results for sensitivity, specificity and accuracy obtained by IMS-ELISA and FC did not show any significant difference (p>0.05). However, when considering laboratory time, cost, ease of operation and the screening of large samples for HLA-B27, the IMS-ELISA was superior to the FC method. We conclude that IMS-ELISA may be used as a fast screening method for HLA B27 detection.
Chou
C T
CT
Division of Allergy-Immunology-Rheumatology, Department of Medicine, Veterans General Hospital, No. 201, Sec. 2, Shih-Pai Road, Shih-Pai, Taipei, Taiwan. ctchou@vghtpe.gov.tw
Tsai
Y F
YF
Liu
J
J
Wei
J C
JC
Liao
T S
TS
Chen
M L
ML
Liu
L Y
LY
D004797:Enzyme-Linked Immunosorbent Assay / Q000191:economics / Q000379:methods*; D005434:Flow Cytometry / Q000191:economics / Q000379:methods; D015796:HLA-B27 Antigen / Q000032:analysis*; D006650:Histocompatibility Testing; D006801:Humans; D018189:Immunomagnetic Separation / Q000191:economics / Q000379:methods*; D008403:Mass Screening / Q000191:economics / Q000379:methods; D016133:Polymerase Chain Reaction; D013167:Spondylitis, Ankylosing / Q000175:diagnosis*
D015796:HLA-B27 Antigen