Flow Cytometric Screening for the <scp>HLA</scp>‐B27 Antigen on Peripheral Blood Lymphocytes

Levering, Wilfried H.B.M.; Sintnicolaas, K.; Wind, Henk; Hooijkaas, Herbert; Gratama, Jan W.

doi:10.1002/0471142956.cy0622s33

Cited by 8 publications

(8 citation statements)

References 29 publications

(44 reference statements)

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“…Notably, mAb 3E12 cross-reactivity on HLA-A2 can be blocked by the addition of anti-HLA-A2-specific mAb clone BB7.2 (results not shown). Interestingly, similar observation was made with a commercial anti-HLA-B27 mAb (clone GS145.2), whose cross-reactivity on HLA-B17, -B37 and -B47 specificities is blocked by the addition of mAb clone FD705 (23,24).…”

Section: Direct If Staining Of Whole Blood Samplessupporting

confidence: 72%

Rapid screening for the detection of HLA‐B57 and HLA‐B58 in prevention of drug hypersensitivity

et al. 2011

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HLA-B57 and HLA-B58 are major histocompatibility class (MHC)-I allotypes that are potentially predictive of important clinical immune phenotypes. HLA-B*5701 is strongly associated with hypersensitivity to the HIV drug abacavir, liver toxicity from the antibiotic flucloxacillin and is a marker for slow progression of HIV AIDS. HLA-B*5801 is associated with hypersensitivity to allopurinol used to treat hyperuricaemia and recurrent gout. Here we describe a monoclonal antibody (mAb) specific for HLA-B57 and HLA-B58 that provides an inexpensive and sensitive screen for these MHC-I allotypes. The usefulness of HLA-B57 screening for prediction of abacavir hypersensitivity was shown in three independent laboratories, including confirmation of the mAb sensitivity and specificity in a cohort of patients enrolled in the PREDICT-1 trial. Our data show that patients who test negative by mAb screening comprise 90%-95% of all individuals in most human populations and require no further human leukocyte antigen (HLA) typing. Patients who test positive by mAb screening should proceed to high-resolution typing to ascertain the presence of HLA-B*5701 or HLA-B*5801. Hence, mAb screening provides a low-cost alternative to high-resolution typing of all patients and lends itself to point-of-care diagnostics and rapid ascertainment of low-risk patients who can begin immediate therapy with abacavir, flucloxacillin or allopurinol.

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Section: Direct If Staining Of Whole Blood Samplessupporting

confidence: 72%

Rapid screening for the detection of HLA‐B57 and HLA‐B58 in prevention of drug hypersensitivity

et al. 2011

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“…The photomultiplier tube voltage was adjusted daily to maintain a constant analysis window using FACSComp software with CaliBRITE beads and HLA‐B27 beads (4, 5). For this study, two different GS145.2‐conjugated antibody lot numbers were used.…”

Section: Methodsmentioning

confidence: 99%

“…The photomultiplier tube window was adjusted daily to maintain a constant analysis window using FACSComp software with CaliBRITE beads (4, 5).…”

Section: Methodsmentioning

confidence: 99%

HLA‐B27 typing: Evaluation of an allele‐specific PCR melting assay and two flow cytometric antigen assays

Seipp

Erali

Wies

et al. 2004

Cytometry Part B Clinical

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Background: Human leukocyte antigen B27 (HLA-B27) is a major histocompatibility complex class 1 molecule that is strongly associated with the disease ankylosing spondylitis. Testing for HLA-B27 is of diagnostic value because 90% of patients with ankylosing spondylitis have the B27 antigen. Two commonly used HLA-B27 flow cytometric assays are commercially available.Methods: An allele-specific polymerase chain reaction (PCR) melting assay for HLA-B27 was compared with two available antigen assays on 371 clinical samples. The accuracy of the assays was measured by receiver operating characteristic analysis using the PCR method and sequencing as the reference standard.Results: When PCR results were compared with those of the antigen assays, complete concordance was observed except for five discrepant results that were resolved by sequence analysis. Using DNA sequencing as the gold standard, the sensitivity and specificity of PCR were 99.6 and 100.0, those of the best single antigen assay were 98.2 and 97.6, and those of a reflex combination of both antigen assays were 98.8 and 97.6.Conclusions: The allele-specific PCR melting assay for HLA-B27 genotyping is easy to perform and has better sensitivity and specificity than antigen assays. The performance of the two flow cytometric antigen assays depends on the antibody used and the positive cutoff values assigned.

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“…For flow cytometry of the parameters HLA-B27, CD-45 and CD-34 + lymphocytes, studies were performed and showed inter-laboratory variation [38,[71][72][73][74]. The competition with PCR methodology hampered further efforts.…”

Section: Hemocytometry and Flow Cytometrymentioning

confidence: 99%

The quest for equivalence of test results: the pilgrimage of the Dutch Calibration 2.000 program for metrological traceability

Jansen

Cobbaert

Weykamp

et al. 2018

Clinical Chemistry and Laboratory Medicine (CCLM)

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Calibration 2.000 was initiated 20 years ago for standardization and harmonization of medical tests. The program also intended to evaluate adequate implementation of the In Vitro Diagnostics (IVD) 98/79/EC directive, in order to ensure that medical tests are fit-for-clinical purpose. The Calibration 2.000 initiative led to ongoing verification of test standardization and harmonization in the Netherlands using commutable external quality assessment (EQA)-tools and a type 1 EQA-design, where feasible. National support was guaranteed by involving all laboratory professionals as well as laboratory technicians responsible for EQA and quality officers. A category 1 EQA-system for general chemistry analytes, harmonizers for specific analytes like hGH and IGF-1, and commutable materials for other EQA-sections have been developed and structurally introduced in the EQA-schemes. The type 1 EQA-design facilitates the dialogue between individual specialists in laboratory medicine and the IVD-industry to reduce lot-to-lot variation and to improve standardization. In such a way, Calibration 2.000 sheds light on the metrological traceability challenges that we are facing and helps the laboratory community to get the issues on the table and resolved. The need for commutable trueness verifiers and/or harmonizers for other medical tests is now seen as paramount. Much knowledge is present in the Netherlands and for general chemistry, humoral immunology and protein chemistry, a few endocrinology tests, and various therapeutic drug monitoring (TDM) tests, commutable materials are available. Also the multi sample evaluation scoring system (MUSE) and the category 1 EQA-design offer many possibilities for permanent education of laboratory professionals to further improve the between and within laboratory variation and the test equivalence.

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Flow Cytometric Screening for the HLA‐B27 Antigen on Peripheral Blood Lymphocytes

Cited by 8 publications

References 29 publications

Rapid screening for the detection of HLA‐B57 and HLA‐B58 in prevention of drug hypersensitivity

Rapid screening for the detection of HLA‐B57 and HLA‐B58 in prevention of drug hypersensitivity

HLA‐B27 typing: Evaluation of an allele‐specific PCR melting assay and two flow cytometric antigen assays

The quest for equivalence of test results: the pilgrimage of the Dutch Calibration 2.000 program for metrological traceability

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