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HLA-B27 testing in Thai patients using the PCR-SSP technique
O Nathalang1, S Tantimavanich, K Nillakupt
1Department of Pathology, Phramongkutklao College of Medicine, Bangkok, Thailand. oytipnti@hotmail.com
Abstract:
We develop the HLA-B27 test kit using the PCR-SSP technique. Five hundred forty blood samples were tested for HLA-B27 by microlymphocytotoxicity test (LCT) and PCR-SSP. It was found that 127 (23.5%) and 134 (24.8%) of these samples were positive for HLA-B27 by LCT and PCR-SSP, respectively. The sensitivity and specificity of the PCR-SSP were 94.8 and 100%, respectively, when using LCT as the standard method. The PCR-SSP positive predictive value was 100%, negative predictive value was 98.3%, and a concordance rate of 98.7%. This study shows that the PCR-SSP is simple, convenient, and a more cost-effective in-house test kit.
Insights
A new Polymerase Chain Reaction Sequence Specific Primer (PCR-SSP) test kit for HLA-B27 was developed. This cost-effective kit demonstrates high accuracy, making it a valuable tool for clinical diagnostics.
Area of Science:
- Immunogenetics
- Molecular Diagnostics
Background:
- The Human Leukocyte Antigen B27 (HLA-B27) gene is associated with various autoimmune diseases.
- Accurate and efficient detection of HLA-B27 is crucial for diagnosis and patient management.
Purpose of the Study:
- To develop and validate a Polymerase Chain Reaction Sequence Specific Primer (PCR-SSP) based test kit for the detection of HLA-B27.
- To evaluate the performance of the developed PCR-SSP kit compared to the gold standard microlymphocytotoxicity test (LCT).
Main Methods:
- Development of an in-house HLA-B27 test kit utilizing the PCR-SSP technique.
- Testing of 540 blood samples using both the newly developed PCR-SSP kit and the established LCT method.
Main Results:
- The PCR-SSP method identified 134 (24.8%) HLA-B27 positive samples, compared to 127 (23.5%) by LCT.
- The PCR-SSP kit demonstrated high sensitivity (94.8%) and perfect specificity (100%) against the LCT method.
- Excellent concordance (98.7%), positive predictive value (100%), and negative predictive value (98.3%) were observed for the PCR-SSP test.
Conclusions:
- The developed PCR-SSP test kit is a simple, convenient, and cost-effective method for HLA-B27 detection.
- PCR-SSP offers a reliable alternative for in-house HLA-B27 testing in clinical laboratories.
- This validated kit can aid in the timely diagnosis of HLA-B27 associated conditions.
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