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Establishment and validation of analytical reference panels for the standardization of quantitative BCR-ABL1 measurements on the international scale

Establishment and validation of analytical reference panels for the standardization of quantitative BCR-ABL1 measurements on the international scale
Establishment and validation of analytical reference panels for the standardization of quantitative BCR-ABL1 measurements on the international scale
Background: Current guidelines for managing Philadelphia-positive chronic myeloid leukemia include monitoring the expression of the BCR-ABL1 (breakpoint cluster region/c-abl oncogene 1, non-receptor tyrosine kinase) fusion gene by quantitative reverse-transcription PCR (RT-qPCR). Our goal was to establish and validate reference panels to mitigate the interlaboratory imprecision of quantitative BCR-ABL1 measurements and to facilitate global standardization on the international scale (IS).

Methods: Four-level secondary reference panels were manufactured under controlled and validated processes with synthetic Armored RNA Quant molecules (Asuragen) calibrated to reference standards from the WHO and the NIST. Performance was evaluated in IS reference laboratories and with non–IS-standardized RT-qPCR methods.

Results: For most methods, percent ratios for BCR-ABL1 e13a2 and e14a2 relative to ABL1 or BCR were robust at 4 different levels and linear over 3 logarithms, from 10% to 0.01% on the IS. The intraassay and interassay imprecision was <2-fold overall. Performance was stable across 3 consecutive lots, in multiple laboratories, and over a period of 18 months to date. International field trials demonstrated the commutability of the reagents and their accurate alignment to the IS within the intra- and interlaboratory imprecision values of IS-standardized methods.

Conclusions: The synthetic calibrator panels are robust, reproducibly manufactured, analytically calibrated to the WHO primary standards, and compatible with most BCR-ABL1 RT-qPCR assay designs. The broad availability of secondary reference reagents will further facilitate interlaboratory comparative studies and independent quality-assessment programs, which are of paramount importance for worldwide standardization of BCR-ABL1 monitoring results and the optimization of current and new therapeutic approaches for chronic myeloid leukemia.

938-948
White, H. E.
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Hedges, J.
e7dcb103-5259-44bc-9f15-bf856a2a1d03
Bendit, I.
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Branford, S.
b3cb17f7-3c04-47be-9b9e-2e9e1f102199
Colomer, D.
fce349a2-3c24-4173-8095-6aca7903719a
Hochhaus, A.
4c0b9da4-adfa-4253-8af7-78cec9e9a24f
Hughes, T.
e7ab9d6a-57d7-45d3-9a1b-bc00df50432e
Kamel-Reid, S.
8c510e63-b007-4449-9c2b-926c3049124e
Kim, D.-W.
668a5867-f060-445c-a52a-777fc8b5d55d
Modur, V.
f6cac275-bd5e-4f30-8aa5-11a61c8a69b1
Muller, M C.
3ab89501-cd26-4fd6-aa2f-b23f5cb58c3b
Pagnano, K.B.
f9a3af9c-6fdc-4889-bb37-76db07ce3dbf
Pane, F.
f289c8fb-c198-41a7-8e08-720ef098c681
Radich, J.
c10289d7-1677-4ea8-a9c2-069462afff0e
Cross, N.C.P.
f87650da-b908-4a34-b31b-d62c5f186fe4
Labourier, E.
718bb3ad-e4fe-4fe5-b2e1-7329a4a306eb
White, H. E.
2181c0b9-fc3b-407e-95eb-3510524603e5
Hedges, J.
e7dcb103-5259-44bc-9f15-bf856a2a1d03
Bendit, I.
5fc5eca3-1035-4453-b073-02c3dd985f93
Branford, S.
b3cb17f7-3c04-47be-9b9e-2e9e1f102199
Colomer, D.
fce349a2-3c24-4173-8095-6aca7903719a
Hochhaus, A.
4c0b9da4-adfa-4253-8af7-78cec9e9a24f
Hughes, T.
e7ab9d6a-57d7-45d3-9a1b-bc00df50432e
Kamel-Reid, S.
8c510e63-b007-4449-9c2b-926c3049124e
Kim, D.-W.
668a5867-f060-445c-a52a-777fc8b5d55d
Modur, V.
f6cac275-bd5e-4f30-8aa5-11a61c8a69b1
Muller, M C.
3ab89501-cd26-4fd6-aa2f-b23f5cb58c3b
Pagnano, K.B.
f9a3af9c-6fdc-4889-bb37-76db07ce3dbf
Pane, F.
f289c8fb-c198-41a7-8e08-720ef098c681
Radich, J.
c10289d7-1677-4ea8-a9c2-069462afff0e
Cross, N.C.P.
f87650da-b908-4a34-b31b-d62c5f186fe4
Labourier, E.
718bb3ad-e4fe-4fe5-b2e1-7329a4a306eb

White, H. E., Hedges, J., Bendit, I., Branford, S., Colomer, D., Hochhaus, A., Hughes, T., Kamel-Reid, S., Kim, D.-W., Modur, V., Muller, M C., Pagnano, K.B., Pane, F., Radich, J., Cross, N.C.P. and Labourier, E. (2013) Establishment and validation of analytical reference panels for the standardization of quantitative BCR-ABL1 measurements on the international scale. Clinical Chemistry, 59 (6), 938-948. (doi:10.1373/clinchem.2012.196477).

Record type: Article

Abstract

Background: Current guidelines for managing Philadelphia-positive chronic myeloid leukemia include monitoring the expression of the BCR-ABL1 (breakpoint cluster region/c-abl oncogene 1, non-receptor tyrosine kinase) fusion gene by quantitative reverse-transcription PCR (RT-qPCR). Our goal was to establish and validate reference panels to mitigate the interlaboratory imprecision of quantitative BCR-ABL1 measurements and to facilitate global standardization on the international scale (IS).

Methods: Four-level secondary reference panels were manufactured under controlled and validated processes with synthetic Armored RNA Quant molecules (Asuragen) calibrated to reference standards from the WHO and the NIST. Performance was evaluated in IS reference laboratories and with non–IS-standardized RT-qPCR methods.

Results: For most methods, percent ratios for BCR-ABL1 e13a2 and e14a2 relative to ABL1 or BCR were robust at 4 different levels and linear over 3 logarithms, from 10% to 0.01% on the IS. The intraassay and interassay imprecision was <2-fold overall. Performance was stable across 3 consecutive lots, in multiple laboratories, and over a period of 18 months to date. International field trials demonstrated the commutability of the reagents and their accurate alignment to the IS within the intra- and interlaboratory imprecision values of IS-standardized methods.

Conclusions: The synthetic calibrator panels are robust, reproducibly manufactured, analytically calibrated to the WHO primary standards, and compatible with most BCR-ABL1 RT-qPCR assay designs. The broad availability of secondary reference reagents will further facilitate interlaboratory comparative studies and independent quality-assessment programs, which are of paramount importance for worldwide standardization of BCR-ABL1 monitoring results and the optimization of current and new therapeutic approaches for chronic myeloid leukemia.

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More information

Published date: 7 March 2013
Organisations: Human Development & Health

Identifiers

Local EPrints ID: 349804
URI: http://eprints.soton.ac.uk/id/eprint/349804
PURE UUID: 3cd173a9-1f40-47c8-b2ca-919fc68bd889
ORCID for N.C.P. Cross: ORCID iD orcid.org/0000-0001-5481-2555

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Date deposited: 11 Mar 2013 14:48
Last modified: 09 Jan 2022 03:08

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Contributors

Author: H. E. White
Author: J. Hedges
Author: I. Bendit
Author: S. Branford
Author: D. Colomer
Author: A. Hochhaus
Author: T. Hughes
Author: S. Kamel-Reid
Author: D.-W. Kim
Author: V. Modur
Author: M C. Muller
Author: K.B. Pagnano
Author: F. Pane
Author: J. Radich
Author: N.C.P. Cross ORCID iD
Author: E. Labourier

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