Establishment and Validation of Analytical Reference Panels for the Standardization of Quantitative BCR-ABL1 Measurements on the International Scale

White, Helen; Hedges, John; Bendit, Israel; Branford, Susan; Colomer, Dolors; Hochhaus, Andreas; Hughes, Timothy P.; Kamel‐Reid, Suzanne; Kim, Dong Wook; Modur, Vijay; Müller, Martin C.; Pagnano, Kátia Bórgia Barbosa; Pane, Fabrizio; Radich, Jerry; Cross, Nicholas C. P.; Labourier, Emmanuel

doi:10.1373/clinchem.2012.196477

Cited by 46 publications

(70 citation statements)

References 28 publications

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“…[2,5] Although the 30 samples from IRIS upon which the IS baseline level was defined have been exhausted, laboratories are able to calibrate their individual BCR-ABL1 RQ-PCR assays to the IS by either of two methods. The first method requires an exchange of a series of samples (20)(21)(22)(23)(24)(25)(26)(27)(28)(29)(30), spanning the entire analytic measurement range) with a reference laboratory that has already been calibrated to the IS and that maintains strict quality control standards for BCR-ABL1 RQ-PCR. [13,15,18] Following the sample exchange, a laboratory-specific conversion factor (CF) is derived using linear regression and bias analyses.…”

Section: Rationale For Development Of the International Scalementioning

confidence: 99%

“…Several of these secondary BCR-ABL1 reference standards are now commercially available, separately and/or packaged into BCR-ABL1 testing kits, although they are not yet FDA approved. [27] The ideal secondary reference standard must be robustly calibrated (on the IS) to the WHO primary reference standard and must cover the entire broad range of expected BCR-ABL1 levels; it must also be composed of material that undergoes all steps of the RQ-PCR process (including RNA extraction) and be stable from time-dependent degradation, available in large quantities, and affordable. With broad availability of secondary reference standards, laboratories will be able to routinely monitor assay accuracy, precision, and time-dependent drift and, thus, may implement the same quality control procedures as they would for any other laboratory analyte in the clinical diagnostics arena.…”

Section: Rationale For Development Of the International Scalementioning

confidence: 99%

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Measurement ofBCR-ABL1transcripts on the International Scale in the United States: current status and best practices

Arora

Press

2016

Leukemia & Lymphoma

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Section: Rationale For Development Of the International Scalementioning

confidence: 99%

Section: Rationale For Development Of the International Scalementioning

confidence: 99%

Measurement ofBCR-ABL1transcripts on the International Scale in the United States: current status and best practices

Arora

Press

2016

Leukemia & Lymphoma

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“…They are used for quality assurance, interlaboratory proficiency testing, and preparation of calibrators or materials to convert quantitative PCR data to the international scale 1 . RMs are usually based on fragments of genomic DNA, cDNA molecules or amplicons of patients suffering from a genetic disease, immortalized cell lines with specific genetic alterations 2,3 , recombinant molecules obtained by targeted mutagenesis 4 , and synthetically prepared genes 5 .…”

Section: Introductionmentioning

confidence: 99%

Preparing compound heterozygous reference material using gene synthesis technology: a model of thrombophilic mutations

Beránek¹,

Drastíková²,

Dulíček³

et al. 2014

BIOMED PAP

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Aims. The aim of our study is to present a novel approach for preparing a compound heterozygous reference material (hetRM) using gene synthesis technology with inverted insertion of wild-type and mutant fragments into a single cloning vector. Factor II (G20210A) and Factor V (G1691A Leiden) gene mutations were used as an experimental model. Methods. During the gene synthesis, DNA fragments were aligned in the following order: G1691 FV wild-type forward strain, G20210 FII wild-type forward strain, 1691A FV mutant reverse strain, 20210A FII mutant reverse strain. The complete chain was inserted into a pIDT SMART cloning vector and amplified in an E. coli competent strain. For assessing hetRM characteristics and commutability, we used real-time PCR with subsequent melting curve analysis, real-time PCR with hydrolysis probes, allele-specific amplification, reverse hybridization, and dideoxynucleotide DNA sequencing. Result. All five methods yielded concordant results of DNA analysis of the hetRM. Differences in real-time PCR cycle threshold values after six-months of storage at -80 °C were not statistically significant from those obtained from freshly prepared hetRM aliquots, which is a good indication of their stability. Conclusion. By applying the procedures of gene synthesis and cloning technology, we prepared and verified a model genetic reference material for FII G20210A and FV G1691A testing with a compound heterozygous genotype. The hetRM was stable, commutable, and available in large quantities and in a wide concentration range.

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“…12, 13 The WHO primary standards consist of a 4-level panel of e14a2-positive lyophilized cell line dilutions. Each level has an assigned IS reference value, the mean IS % ratio that was obtained by repeated testing of each sample level in expert IS-standardized laboratories during the establishment of these materials.…”

mentioning

confidence: 99%

“…Each level has an assigned IS reference value, the mean IS % ratio that was obtained by repeated testing of each sample level in expert IS-standardized laboratories during the establishment of these materials. 13 Approximately 3500 primary reference panels were initially manufactured. To ensure the long-term supply of stable, homogeneous, commutable, and traceable reference standards to hundreds of clinical laboratories worldwide, secondary reference panels were developed and validated.…”

mentioning

confidence: 99%

Conversion, Correction, and International Scale Standardization: Results From a Multicenter External Quality Assessment Study forBCR-ABL1Testing

Griffiths

Patton

Grossi

et al. 2015

Archives of Pathology & Laboratory Medicine

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Context.-Monitoring BCR-ABL1 expression levels relative to clinically validated response criteria on the International Scale (IS) is vital in the optimal management of patients with chronic myeloid leukemia, yet significant variability remains across laboratories worldwide.Objective.-To assess method performance, interlaboratory precision, and different IS standardization modalities in representative laboratories performing routine BCR-ABL1 testing.Design.-Fifteen blinded test specimens with 5-level nominal BCR-ABL1 to ABL1 IS percentage ratios ranging from 5% to 0.0005% and 4-level secondary IS reference panels, the ARQ IS Calibrator Panels, were tested by relative quantitative polymerase chain reaction in 15 laboratories in 5 countries. Both raw and IS percentage ratios calculated by using local conversion factors (CFs) or analytic correction parameters (CPs) were collected and analyzed.Results.-A total of 670 valid positive results were generated. BCR-ABL1 detection was associated with variable ABL1 quality metric passing rates (P , .001) and reached at least 0.01% in 13 laboratories. Intralaboratory precision was within 2.5-fold for all sample levels combined with a relative mean difference greater than 5-fold across laboratories. International Scale accuracy was increased by using both the CF and CP standardization methods. Classification agreement for major molecular response status was 90% after CF conversion and 93% after CP correction, with precision improved by 3-fold for the CP method.Conclusions.-Despite preanalytic and analytic differences between laboratories, conversion and correction are effective IS standardization methods. Validated secondary reference materials can facilitate global diffusion of the IS without the need to perform sample exchange and improve the accuracy and precision of BCR-ABL1 quantitative measurements, including at low levels of residual disease.

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Establishment and Validation of Analytical Reference Panels for the Standardization of Quantitative BCR-ABL1 Measurements on the International Scale

Cited by 46 publications

References 28 publications

Measurement ofBCR-ABL1transcripts on the International Scale in the United States: current status and best practices

Measurement ofBCR-ABL1transcripts on the International Scale in the United States: current status and best practices

Preparing compound heterozygous reference material using gene synthesis technology: a model of thrombophilic mutations

Conversion, Correction, and International Scale Standardization: Results From a Multicenter External Quality Assessment Study forBCR-ABL1Testing

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