Evaluation of Two Commercial HLA-B27 Real-Time PCR Kits
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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 HLA-B27 real-time PCR kit (Bioneer, Korea) with TaqMan probes and the Real-QTM 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 realtime 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.
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 HLA-B27 real-time PCR kit (Bioneer, Korea) with TaqMan probes and the Real-QTM 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 realtime 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.
참고문헌 (12)
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