Comparison of HLA-B27 typing methods -PCR-SSP, microlymphocytotoxicity, and flow cytometry.
- Author:
Jun Soo BAE
1
;
Young Ree KIM
;
Hyun Il CHOI
;
Yun Jung CHO
Author Information
1. Department of Clinical Pathology, Korea University Medical College. eqchokum@ns.kumc.or.kr
- Publication Type:Original Article
- Keywords:
HLA-B27;
HLA-B7;
PCR-SSP;
microlymphocytotoxicity;
flow cytometry
- MeSH:
Flow Cytometry*;
HLA-B Antigens;
HLA-B27 Antigen*;
HLA-B7 Antigen
- From:Korean Journal of Clinical Pathology
2000;20(2):198-203
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
BACKGROUND: HLA-B27 typing has long been performed by the microlymphocytotoxicity method(MCT) but the flow cytometry method(FCM) was introduced several years ago. False positive results due to the HLA-B7 cross reactive groups(CREG) were the main drawback of the serologic method. The authors performed polymerase chain reaction-sequence specific primer(PCR-SSP) test for HLA-B27 to compare the results with serologic methods. METHODS: PCR-SSP test for HLA-B27 was performed on four hundred forty one samples. Three hundred twenty eight samples were tested by MCT and one hundred thirteen samples by FCM. PCR-SSP for HLA-B27 subtyping or Amplification Refractory Mutation System-PCR(ARMS-PCR) for HLA-B typing was performed on twenty four discrepant samples. RESULTS: The concordance rate between MCT and PCR-SSP was 92.9%(305/328) and the concordance rate between FCM and PCR-SSP was 99.1%(112/113). Twenty four(5.4%) out of four hundred forty one samples showed discrepancy between serologic methods and PCR-SSP method. Fourteen out of one hundred MCT positive samples and only one out of forty FCM positive samples showed negative by PCR-SSP. Nine samples showed PCR-SSP positive and MCT negative. CONCLUSIONS: The false positive rate of MCT was quite high and there were some false positive and negative results by PCR-SSP, too. From the above findings, we suggest that FCM is the most accurate method for HLA-B27 typing in those laboratory equipped with flow cytometry.
