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Sequence polymorphism systems for quantitative real-time polymerase chain reaction to characterize hematopoietic chimerism-high informativity and sensitivity as well as excellent reproducibility and precision of measurement.
Lab Hematol. 2007; 13(3):73-84.LH

Abstract

Sequence polymorphisms (SPs) can serve as genetic markers for quantitative polymerase chain reactions (qPCR) for chimerism analysis, providing a significantly higher sensitivity compared to short tandem repeat PCR. In this study, a panel of 29 selected markers was evaluated in 317 patients with leukemia and myelodysplastic syndrome, who received allogeneic stem cell transplantation. In total, 5415 posttransplantation samples were analyzed. Recipient genotype discrimination was possible in 96% with a mean number of 2.5 (1-7) informative markers per recipient/donor pair. Marker specific standard dilution series from volunteers' DNA served as standard for quantification of chimerism. Sensitivity of the method was < or =1 x 10-3 (0.1% of recipient cells) in 83.3% of the assays. By this method, it was possible to very accurately detect autologous signals in the range from 0% to 0.5% (95% confidence interval [CI] +/-0.2), from 0.5% to 1% (95% CI +/-0.4), from 1% to 2% (95% CI +/-0.6) and from 2% to 5% (95% CI +/-1.2). Reproducibility of the quantified autologous signals was independent from the amount of DNA. This is the first report on a SP-based chimerism system allowing for the performance of chimerism analyses for virtually all patients with high sensitivity, excellent reproducibility, and precision of measurement.

Authors+Show Affiliations

J. W. Goethe University Children's Hospital of Frankfurt, Frankfurt, Germany.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Evaluation Study
Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

17984038

Citation

Willasch, Andre, et al. "Sequence Polymorphism Systems for Quantitative Real-time Polymerase Chain Reaction to Characterize Hematopoietic Chimerism-high Informativity and Sensitivity as Well as Excellent Reproducibility and Precision of Measurement." Laboratory Hematology : Official Publication of the International Society for Laboratory Hematology, vol. 13, no. 3, 2007, pp. 73-84.
Willasch A, Schneider G, Reincke BS, et al. Sequence polymorphism systems for quantitative real-time polymerase chain reaction to characterize hematopoietic chimerism-high informativity and sensitivity as well as excellent reproducibility and precision of measurement. Lab Hematol. 2007;13(3):73-84.
Willasch, A., Schneider, G., Reincke, B. S., Shayegi, N., Kreyenberg, H., Kuci, S., Weber, G., Van Der Reijden, B., Niethammer, D., Klingebiel, T., & Bader, P. (2007). Sequence polymorphism systems for quantitative real-time polymerase chain reaction to characterize hematopoietic chimerism-high informativity and sensitivity as well as excellent reproducibility and precision of measurement. Laboratory Hematology : Official Publication of the International Society for Laboratory Hematology, 13(3), 73-84.
Willasch A, et al. Sequence Polymorphism Systems for Quantitative Real-time Polymerase Chain Reaction to Characterize Hematopoietic Chimerism-high Informativity and Sensitivity as Well as Excellent Reproducibility and Precision of Measurement. Lab Hematol. 2007;13(3):73-84. PubMed PMID: 17984038.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Sequence polymorphism systems for quantitative real-time polymerase chain reaction to characterize hematopoietic chimerism-high informativity and sensitivity as well as excellent reproducibility and precision of measurement. AU - Willasch,Andre, AU - Schneider,Gaby, AU - Reincke,Britta Sofia, AU - Shayegi,Nona, AU - Kreyenberg,Hermann, AU - Kuci,Selim, AU - Weber,Gerrit, AU - Van Der Reijden,Bert, AU - Niethammer,Dietrich, AU - Klingebiel,Thomas, AU - Bader,Peter, PY - 2007/11/7/pubmed PY - 2008/1/16/medline PY - 2007/11/7/entrez SP - 73 EP - 84 JF - Laboratory hematology : official publication of the International Society for Laboratory Hematology JO - Lab Hematol VL - 13 IS - 3 N2 - Sequence polymorphisms (SPs) can serve as genetic markers for quantitative polymerase chain reactions (qPCR) for chimerism analysis, providing a significantly higher sensitivity compared to short tandem repeat PCR. In this study, a panel of 29 selected markers was evaluated in 317 patients with leukemia and myelodysplastic syndrome, who received allogeneic stem cell transplantation. In total, 5415 posttransplantation samples were analyzed. Recipient genotype discrimination was possible in 96% with a mean number of 2.5 (1-7) informative markers per recipient/donor pair. Marker specific standard dilution series from volunteers' DNA served as standard for quantification of chimerism. Sensitivity of the method was < or =1 x 10-3 (0.1% of recipient cells) in 83.3% of the assays. By this method, it was possible to very accurately detect autologous signals in the range from 0% to 0.5% (95% confidence interval [CI] +/-0.2), from 0.5% to 1% (95% CI +/-0.4), from 1% to 2% (95% CI +/-0.6) and from 2% to 5% (95% CI +/-1.2). Reproducibility of the quantified autologous signals was independent from the amount of DNA. This is the first report on a SP-based chimerism system allowing for the performance of chimerism analyses for virtually all patients with high sensitivity, excellent reproducibility, and precision of measurement. SN - 1080-2924 UR - https://www.unboundmedicine.com/medline/citation/17984038/Sequence_polymorphism_systems_for_quantitative_real_time_polymerase_chain_reaction_to_characterize_hematopoietic_chimerism_high_informativity_and_sensitivity_as_well_as_excellent_reproducibility_and_precision_of_measurement_ L2 - https://ClinicalTrials.gov/search/term=17984038 [PUBMED-IDS] DB - PRIME DP - Unbound Medicine ER -