A new highly sensitive real-time quantitative-PCR method for detection of BCR-ABL1 to monitor minimal residual disease in chronic myeloid leukemia after discontinuation of imatinib

Hiroaki Kitamura, Yoko Tabe, Tomohiko Ai, Koji Tsuchiya, Maiko Yuri, Shigeki Misawa, Takashi Horii, Atsushi Kawaguchi, Akimichi Ohsaka, Shinya Kimura

Research output: Contribution to journalArticle

Abstract

Tyrosine kinase inhibitors (TKIs) targeting the BCR-ABL1 fusion protein, encoded by the Philadelphia chromosome, have drastically improved the outcomes for patients with chronic myeloid leukemia (CML). Although several real-time quantitative polymerase chain reaction (RQ-PCR) kits for the detection of BCR-ABL1 transcripts are commercially available, their accuracy and efficiency in laboratory practice require reevaluation. We have developed a new in-house RQ-PCR method to detect minimal residual disease (MRD) in CML cases. MRD was analyzed in 102 patients with CML from the DOMEST study, a clinical trial to study the rationale for imatinib mesylate discontinuation in Japan. The BCR-ABL1/ABL1 ratio was evaluated using the international standard (IS) ratio, where IS < 0.1% was defined as a major molecular response. At enrollment, BCR-ABL1 transcripts were undetectable in all samples using a widely-applied RQ-PCR method performed in the commercial laboratory, BML (BML Inc., Tokyo, Japan); however, the in-house method detected the BCR-ABL1 transcripts in five samples (5%) (mean IS ratio: 0.0062 ± 0.0010%). After discontinuation of imatinib, BCR-ABL1 transcripts were detected using the in-house RQ-PCR in 21 patients (21%) that were not positive using the BML method. Nineteen samples were also tested using a commercially available RQ-PCR assay kit with a detection limit of IS ratio, 0.0032 (ODK-1201, Otsuka Pharmaceutical Co., Tokyo, Japan). This method detected low levels of BCR-ABL1 transcripts in 14 samples (74%), but scored negative for five samples (26%) that were positive using the in-house method. From the perspective of the in-house RQ-PCR method, number of patients confirmed loss of MMR was 4. These data suggest that our new in-house RQ-PCR method is effective for monitoring MRD in CML.

Original languageEnglish (US)
Article numbere0207170
JournalPLoS ONE
Volume14
Issue number3
DOIs
StatePublished - Mar 1 2019
Externally publishedYes

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myeloid leukemia
Polymerase chain reaction
Residual Neoplasm
Leukemia, Myelogenous, Chronic, BCR-ABL Positive
chronic diseases
Real-Time Polymerase Chain Reaction
quantitative polymerase chain reaction
monitoring
Japan
Tokyo
methodology
sampling
Philadelphia Chromosome
Chromosomes
Imatinib Mesylate
Protein-Tyrosine Kinases
Assays
Fusion reactions
tyrosine
Limit of Detection

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

A new highly sensitive real-time quantitative-PCR method for detection of BCR-ABL1 to monitor minimal residual disease in chronic myeloid leukemia after discontinuation of imatinib. / Kitamura, Hiroaki; Tabe, Yoko; Ai, Tomohiko; Tsuchiya, Koji; Yuri, Maiko; Misawa, Shigeki; Horii, Takashi; Kawaguchi, Atsushi; Ohsaka, Akimichi; Kimura, Shinya.

In: PLoS ONE, Vol. 14, No. 3, e0207170, 01.03.2019.

Research output: Contribution to journalArticle

Kitamura, Hiroaki ; Tabe, Yoko ; Ai, Tomohiko ; Tsuchiya, Koji ; Yuri, Maiko ; Misawa, Shigeki ; Horii, Takashi ; Kawaguchi, Atsushi ; Ohsaka, Akimichi ; Kimura, Shinya. / A new highly sensitive real-time quantitative-PCR method for detection of BCR-ABL1 to monitor minimal residual disease in chronic myeloid leukemia after discontinuation of imatinib. In: PLoS ONE. 2019 ; Vol. 14, No. 3.
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abstract = "Tyrosine kinase inhibitors (TKIs) targeting the BCR-ABL1 fusion protein, encoded by the Philadelphia chromosome, have drastically improved the outcomes for patients with chronic myeloid leukemia (CML). Although several real-time quantitative polymerase chain reaction (RQ-PCR) kits for the detection of BCR-ABL1 transcripts are commercially available, their accuracy and efficiency in laboratory practice require reevaluation. We have developed a new in-house RQ-PCR method to detect minimal residual disease (MRD) in CML cases. MRD was analyzed in 102 patients with CML from the DOMEST study, a clinical trial to study the rationale for imatinib mesylate discontinuation in Japan. The BCR-ABL1/ABL1 ratio was evaluated using the international standard (IS) ratio, where IS < 0.1{\%} was defined as a major molecular response. At enrollment, BCR-ABL1 transcripts were undetectable in all samples using a widely-applied RQ-PCR method performed in the commercial laboratory, BML (BML Inc., Tokyo, Japan); however, the in-house method detected the BCR-ABL1 transcripts in five samples (5{\%}) (mean IS ratio: 0.0062 ± 0.0010{\%}). After discontinuation of imatinib, BCR-ABL1 transcripts were detected using the in-house RQ-PCR in 21 patients (21{\%}) that were not positive using the BML method. Nineteen samples were also tested using a commercially available RQ-PCR assay kit with a detection limit of IS ratio, 0.0032 (ODK-1201, Otsuka Pharmaceutical Co., Tokyo, Japan). This method detected low levels of BCR-ABL1 transcripts in 14 samples (74{\%}), but scored negative for five samples (26{\%}) that were positive using the in-house method. From the perspective of the in-house RQ-PCR method, number of patients confirmed loss of MMR was 4. These data suggest that our new in-house RQ-PCR method is effective for monitoring MRD in CML.",
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AU - Tsuchiya, Koji

AU - Yuri, Maiko

AU - Misawa, Shigeki

AU - Horii, Takashi

AU - Kawaguchi, Atsushi

AU - Ohsaka, Akimichi

AU - Kimura, Shinya

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