ViroGene HLA B27 QPCR Kit

The human leukocyte antigen HLA-B27 is strongly associated with spondyloarthropathies (SpA), a group of inflammatory rheumatic diseases including ankylosing spondylitis (AS). HLA-B27 is found in 90–95% of AS patients. It is also found in a lower proportion of patients with reactive arthritis and some forms of psoriatic arthritis (PsA). Twenty-four HLA-B27 subtypes have been detected and differ only by a small number of nucleotide substitutions within exons 2 and 3 of the HLA-B27 gene. Although the exact mechanism determining disease susceptibility is still unknown, testing for HLA-B27 is a valuable tool for the diagnosis of AS and SpA

Serological techniques such as the micro lymphocytotoxicity test (MLCT), flow cytometry (FC) and enzyme immunoassays (EIA) are usually used for the routine typing of HLA-B27. However, these techniques require fresh blood samples (viable cells) because they are based on the detection of cell surface structures by antibodies and are thus sensitive to downregulation or conformational changes of the HLA-B27 glycoprotein. Moreover, serological methods lack specificity especially in the presence of antigens that cross-react with HLA-B27, such as HLA-B7. Therefore, several molecular methods that do not require viable cells and are more accurate have been developed for HLA-B27 genotyping. These methods include PCR restriction length polymorphism, polymerase chain reaction with sequence-specific primers (PCR-SSP), hybridization with specific oligonucleotide probes (PCR-SSO), ligation-based typing (LBT) and sequence- based typing (SBT).

The development of real-time PCR was an important technical improvement permitting the detection and quantification of PCR products during thermal cycling. Real-time PCR is a powerful tool for gene expression analysis, genotyping, pathogen detection/ quantification and mutation screening

 

Indication	in vitro diagnostic medical device
Regulatory Status	CE IVD
Intended User	For professional use in laboratories with trained staff
Technology	Real-time PCR
Type of Analysis	Qualitative
Target Sequence	HLA B27 (Exon2 and Exon3)
Analytical Sensitivity (LoD)	Reaches up to 2 ng/µl
Positive Percentage Agreement	100 % (CI95%: 79.95 % – 100 %)
Negative Percentage Agreement	100 % (CI95%: 65.55 % – 100 %)
Overall Percentage Agreement	100 % (CI95%: 85.87 % – 100 %)
Validated Specimen	Whole blood
Storage	-20 ± 5 °C
Validated Extraction Methods	ViroGene DNA Extraction Kit & NA16 MagCore NA Extractor
Instruments	Applied Biosystems 7300 / 7500 Real-Time PCR System AriaMx Real-Time PCR System LightCycler® 2.0 / 480 CFX Connect™ / CFX96™ / Dx Real-Time PCR Detection System LineGene 9600 Plus Mic qPCR Cycler QuantStudio™ 3 / 5 Real-Time PCR System Rotor-Gene 3000/6000/Q* SLAN® Real-Time PCR System StepOne™ / StepOnePlus™ Real-Time PCR System
Required Detection Channels	FAM, HEX/JOE