HLA-B27 typing: Evaluation of an allele-specific PCR melting assay and two flow cytometric antigen assays
نویسندگان
چکیده
منابع مشابه
Validation of allele-specific polymerase chain reaction for DNA typing of HLA-B27.
To find a specific method for HLA-B27 typing for the diagnosis of rheumatic disorders, we extensively tested the single-step B27-specific polymerase chain reaction (PCR) described by Dominguez et al. (Immunogenetics 1992;36:277-82). This method, which relies on specific primer recognition of a sequence in the third exon (unique to the B27-allele), was used for screening of 270 characterized blo...
متن کاملFlow Cytometric Human Leukocyte Antigen-B27 Typing with Stored Samples for Batch Testing
BACKGROUND Flow cytometry (FC) HLA-B27 typing is still used extensively for the diagnosis of spondyloarthropathies. If patient blood samples are stored for a prolonged duration, this testing can be performed in a batch manner, and in-house cellular controls could easily be procured. In this study, we investigated various methods of storing patient blood samples. METHODS WE COMPARED FOUR STORA...
متن کاملHLA-B27 typing by use of flow cytofluorometry.
We describe the use of flow cytofluorometry to type lymphocytes for HLA-B27 antigens. We modified the cytotoxicity test of Terasaki and McClelland [Nature (London) 1964;204:998] for use with a flow cytofluorometer. In a second assay we used a commercially available monoclonal anti-HLA-B27 antibody and indirect immunofluorescence staining of cell surfaces. Although this antibody cross reacts wit...
متن کاملHLA-DQA1 allele typing by nonisotopic PCR-LIS-SSCP.
In the present study we used a simple and reliable method for HLA-DQA1 allele typing based on the single-stranded conformation polymorphism (SSCP) properties of DNA molecules obtained by PCR. The technique consists of PCR amplification of a DNA fragment comprising the second exon of the HLA-DQA1 gene, amplicon denaturation using a low ionic strength solution (LIS), and electrophoresis on a smal...
متن کاملCOMPARISON OF VAL IDITY OF MICROLYMPHOCYTOTOXICITY AND FLOWCYTOMETRY METHODS WITH PCR FOR HL A-B27 ANTIGEN TYPING
The Human Major Histocompatibility Complex (MHC) plays a crucial role in transplantation, transfusion, paternity test and assessment of susceptibility to some diseases associated with HLA-B27. Three of the most fashionable methods for determination of HLA antigens in clinical and research laboratories are microlymphocytotoxicity (MLCT), flowcytometry and polymerase chain reaction (PCR). T...
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ژورنال
عنوان ژورنال: Cytometry Part B: Clinical Cytometry
سال: 2004
ISSN: 1552-4949,1552-4957
DOI: 10.1002/cyto.b.20039