Evaluation of Two Commercial HLA-B27 Real-Time PCR Kits.
10.3343/kjlm.2009.29.6.589
- Author:
Eun Hae CHO
1
;
Sang Gon LEE
;
Jeong Ho SEOK
;
Bo Ya PARK
;
Eun Hee LEE
Author Information
1. Greencross Reference Laboratory, Yongin, Korea. ssea74@yahoo.co.kr
- Publication Type:Original Article ; Evaluation Studies
- Keywords:
Real Time PCR;
HLA-B27;
PCR-SSP
- MeSH:
HLA-B27 Antigen/*analysis;
Histocompatibility Testing/*methods;
Humans;
Organic Chemicals/chemistry;
Polymerase Chain Reaction;
Reagent Kits, Diagnostic;
Transition Temperature
- From:The Korean Journal of Laboratory Medicine
2009;29(6):589-593
- CountryRepublic of Korea
- Language:English
-
Abstract:
BACKGROUND: The standard PCR with sequence-specific primers (SSP) is a widely used method of HLA-B27 typing in clinical practice. The aim of our study was to evaluate 2 Korean HLA-B27 kits with different real-time PCR chemistries. METHODS: To validate the accuracy of real-time PCR kits, we selected 28 HLA-B27-positive samples and 33 HLA-B27-negative samples with a wide range of different HLA-B specificities typed by standard PCR-SSP. The 2 real-time PCR kits used were the AccuPower(R) HLA-B27 real-time PCR kit (Bioneer, Korea) with TaqMan probes and the Real-Q(TM) HLA-B*27 detection kit (BioSewoom, Korea) with SYBR Green I dye for melting curve analysis. RESULTS: All 61 samples typed by PCR-SSP demonstrated a perfect concordance with the 2 real-time PCR assays. It was possible to clearly discriminate between HLA-B27-positive and -negative samples in both real-time assays. CONCLUSIONS: In summary, both real-time PCR assays for HLA-B27 were fast, reliable, well-adapted for routine laboratory testing, and attractive alternatives to the conventional PCR-SSP method.
