Multiplex polymerase chain reaction on FTA cards vs. flow cytometry for B-lymphocyte clonality
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Michael Dictor
Abstract
Background: Two-colour flow cytometry was compared with multiplex PCR with capillary electrophoresis for clonality determination in specific categories of B-cell lymphoma. FTA cards were evaluated for preserving DNA from node imprints and expediting molecular analysis.
Methods: A single-tube multiplex PCR targeted IGH and lymphoma-specific translocations in DNA extracted from 180 frozen lymphoid tissues and DNA bound to FTA cards from 192 fresh tissues and 137 aspirates. PCR results were compared with flow cytometry in the extracted and aspirated samples.
Results: Overall, single-tube multiplex PCR sensitivity was equivalent in the sample groups (intergroup range 79%–91%). False negatives were associated with tumour origin in the follicle centre. Multiplex PCR and flow cytometry were equally sensitive and together detected 98% of B-cell lymphomas. Additional two-tube targeting of IGK suggested an overall molecular sensitivity >90%. False positive (pseudoclonal) single-tube multiplex PCR was associated with necrosis and sparse lymphocytes.
Conclusions: Multiplex PCR using template DNA bound to an FTA card effectively detects B-lymphocyte clonality, obviates DNA extraction and refrigeration, and can be used without diminished sensitivity in fine needle aspirates or node imprints as a replacement for or complement to flow cytometry at any point in the diagnostic work-up.
Clin Chem Lab Med 2007;45:339–45.
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©2007 by Walter de Gruyter Berlin New York
Articles in the same Issue
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Articles in the same Issue
- From human genetic variations to prediction of risks and responses to drugs and the environment
- Nutrigenomics – 2006 update
- How to comprehensively analyse proteins and how this influences nutritional research
- Genotypes, obesity and type 2 diabetes – can genetic information motivate weight loss? A review
- The Gene-Diet Attica Investigation on childhood obesity (GENDAI): overview of the study design
- Polymorphisms in the APOA1/C3/A4/A5 gene cluster and cholesterol responsiveness to dietary change
- Nutri-epigenomics: lifelong remodelling of our epigenomes by nutritional and metabolic factors and beyond
- Emerging role of cathepsin S in obesity and its associated diseases
- Association analysis of hepatitis virus B infection with haplotypes of the TBX21 gene promoter region in the Chinese population
- Multiplex polymerase chain reaction on FTA cards vs. flow cytometry for B-lymphocyte clonality
- Real-time multiplex PCR assay for genotyping of three apolipoprotein E alleles and two choline acetyltransferase alleles with three hybridization probes
- Immunomagnetic CD45 depletion does not improve cytokeratin 20 RT-PCR in colorectal cancer
- Analysis of the components of hypertransaminasemia after liver resection
- Fine characterization of mitral valve glycosaminoglycans and their modification with degenerative disease
- Oxidative stress evaluated using an automated method for hydroperoxide estimation in patients with coronary artery disease
- Secretory phospholipase A2 activity and release kinetics of vascular tissue remodelling biomarkers after coronary artery bypass grafting with and without cardiopulmonary bypass
- Clustered components of the metabolic syndrome and platelet counts in Japanese females
- International Standard for serum vitamin B12 and serum folate: international collaborative study to evaluate a batch of lyophilised serum for B12 and folate content
- Multicentre physiological reference intervals for serum concentrations of immunoglobulins A, G and M, complement C3c and C4 measured with Tina-Quant® reagents systems
- In vivo and in vitro allergy diagnostics: it's time to reappraise the costs
- Experience with post-market surveillance of in-vitro diagnostic medical devices for lay use in Germany
- Evaluation of the high-sensitivity, full-range Olympus CRP OSR6199 application on the Olympus AU640®
- How to improve the teaching of urine microscopy
- In vitro determination of allergen-specific serum IgE. Comparative analysis of three methods
- Efficacy of a new blocker against anti-ruthenium antibody interference in the Elecsys free triiodothyronine assay
- Clinical indications for plasma protein assays: transthyretin (prealbumin) in inflammation and malnutrition: International Federation of Clinical Chemistry and Laboratory Medicine (IFCC): IFCC Scientific Division Committee on Plasma Proteins (C-PP)
- Meeting Report: From human genetic variations to prediction of risks and responses to drugs and the environment
