Analytical Validation of a Highly Sensitive, Multiplexed Chronic Myeloid Leukemia Monitoring System Targeting BCR-ABL1 RNA

Justin T. Brown, Ion J. Beldorth, Walairat Laosinchai-Wolf, Marie E. Fahey, Keri L. Jefferson, Adam K. Ruskin, Jacquelyn J. Roth, Li Cai, Christopher D. Watt, Richard Press, Fei Yang, John B. Hedges, Bernard F. Andruss

Research output: Contribution to journalArticle

Abstract

This study describes the analytical performance of the QuantideX qPCR BCR-ABL IS Kit, the first Food and Drug Administration–cleared assay designed to monitor breakpoint cluster region–Abelson tyrosine-protein kinase 1 (BCR-ABL1) fusion transcripts isolated from peripheral blood specimens from patients with chronic myeloid leukemia. This multiplex real-time quantitative RT-PCR assay amplifies both e13a2 and e14a2 Major BCR-ABL1 transcripts and the reference target ABL1. The test results are provided in international scale (IS) values by incorporating armored RNA-based calibrators that have defined IS values tied directly to the World Health Organization BCR-ABL1 Primary Reference Materials, without the necessity of determining and maintaining conversion factors. For each batch run, the integrated interpretive software evaluates run and specimen quality control metrics (including a sufficient amount of ABL1 control transcripts to ensure a minimal limit of detection) and calculates both molecular response (MR) and %IS values for each specimen. The test has a limit of detection of MR4.7 (0.002%IS) and a linear range from MR0.3 (50%IS) to MR4.7 (0.002%IS) for both Major transcripts. Single-site and multisite precision studies demonstrated a maximum SD of 0.13 MR (30% CV within the assay range between MR0.7 and MR3.7). The performance of this BCR-ABL1 monitoring test meets all of the clinical guideline recommendations for sensitivity and IS reporting for the management of chronic myeloid leukemia patients.

Original languageEnglish (US)
Pages (from-to)718-733
Number of pages16
JournalJournal of Molecular Diagnostics
Volume21
Issue number4
DOIs
StatePublished - Jul 1 2019

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Leukemia, Myelogenous, Chronic, BCR-ABL Positive
Protein-Tyrosine Kinases
RNA
Limit of Detection
Quality Control
Real-Time Polymerase Chain Reaction
Software
Guidelines
Food
Pharmaceutical Preparations

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Molecular Medicine

Cite this

Brown, J. T., Beldorth, I. J., Laosinchai-Wolf, W., Fahey, M. E., Jefferson, K. L., Ruskin, A. K., ... Andruss, B. F. (2019). Analytical Validation of a Highly Sensitive, Multiplexed Chronic Myeloid Leukemia Monitoring System Targeting BCR-ABL1 RNA. Journal of Molecular Diagnostics, 21(4), 718-733. https://doi.org/10.1016/j.jmoldx.2019.03.002

Analytical Validation of a Highly Sensitive, Multiplexed Chronic Myeloid Leukemia Monitoring System Targeting BCR-ABL1 RNA. / Brown, Justin T.; Beldorth, Ion J.; Laosinchai-Wolf, Walairat; Fahey, Marie E.; Jefferson, Keri L.; Ruskin, Adam K.; Roth, Jacquelyn J.; Cai, Li; Watt, Christopher D.; Press, Richard; Yang, Fei; Hedges, John B.; Andruss, Bernard F.

In: Journal of Molecular Diagnostics, Vol. 21, No. 4, 01.07.2019, p. 718-733.

Research output: Contribution to journalArticle

Brown, JT, Beldorth, IJ, Laosinchai-Wolf, W, Fahey, ME, Jefferson, KL, Ruskin, AK, Roth, JJ, Cai, L, Watt, CD, Press, R, Yang, F, Hedges, JB & Andruss, BF 2019, 'Analytical Validation of a Highly Sensitive, Multiplexed Chronic Myeloid Leukemia Monitoring System Targeting BCR-ABL1 RNA', Journal of Molecular Diagnostics, vol. 21, no. 4, pp. 718-733. https://doi.org/10.1016/j.jmoldx.2019.03.002
Brown, Justin T. ; Beldorth, Ion J. ; Laosinchai-Wolf, Walairat ; Fahey, Marie E. ; Jefferson, Keri L. ; Ruskin, Adam K. ; Roth, Jacquelyn J. ; Cai, Li ; Watt, Christopher D. ; Press, Richard ; Yang, Fei ; Hedges, John B. ; Andruss, Bernard F. / Analytical Validation of a Highly Sensitive, Multiplexed Chronic Myeloid Leukemia Monitoring System Targeting BCR-ABL1 RNA. In: Journal of Molecular Diagnostics. 2019 ; Vol. 21, No. 4. pp. 718-733.
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AU - Jefferson, Keri L.

AU - Ruskin, Adam K.

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