title: Validation of a rapid one-step high sensitivity real-time quantitative PCR system for detecting major BCR-ABL1 mRNA on an International Scale.
abstract: BACKGROUND
  Detection and quantitation of BCR-ABL1 transcripts are crucial for managing patients with chronic myeloid leukemia (CML). Although real-time quantitative polymerase chain reaction (RT-qPCR) can be measured on an International Scale (IS), this has not become fully universal. By using a WHO international standard panel established for calibrating secondary standards based on the IS, we have previously developed an RT-qPCR kit, ODK-1201, for quantification of major BCR-ABL1.
  
  
  RESULTS
  In this study, the reliability of kit-specific conversion factor 1.12 was validated by exchanging patients' samples between three local clinical laboratories and a reference laboratory. The mean bias of the local method after IS conversion was 1.6 fold lower than the reference method. The clinically-useful sensitivity of the kit was further evaluated for monitoring patients with deep molecular response. Based on the correlation of the IS values between ODK-1201 and the reference laboratory method, the detection level of the kit was estimated as 0.0032 % BCR-ABL1 (IS).
  
  
  CONCLUSIONS
  ODK-1201 is a highly sensitive one-step RT-qPCR system for detecting BCR-ABL1 on the IS in 2 h after RNA extraction, thus contributing to standardization of molecular monitoring in CML.
authors.affiliation: Department of Hematology, National Hospital Organization Mito Medical Center, 280, Sakuranosato, Ibarakimachi, Higashiibarakigun, Ibaraki, 311-3193 Japan.
authors.forename: Chikashi
authors.initials: C
authors.lastname: Yoshida
authors.affiliation: Hematology, Department of Internal Medicine, Osaka City University Graduate School of Medicine, Osaka, Japan.
authors.forename: Hirohisa
authors.initials: H
authors.lastname: Nakamae
authors.affiliation: Department of Genetic and Molecular Pathology, Centre for Cancer Biology, SA Pathology, Adelaide, Australia.
authors.forename: Linda
authors.initials: L
authors.lastname: Fletcher
authors.affiliation: Diagnostic Division, Otsuka Pharmaceutical, Tokushima, Japan.
authors.forename: Daisuke
authors.initials: D
authors.lastname: Koga
authors.affiliation: Diagnostic Division, Otsuka Pharmaceutical, Tokushima, Japan.
authors.forename: Takayuki
authors.initials: T
authors.lastname: Sogabe
authors.affiliation: Hematology, Department of Internal Medicine, Kinki University School of Medicine, Osaka, Japan.
authors.forename: Itaru
authors.initials: I
authors.lastname: Matsumura
authors.affiliation: Hematology and Oncology, Osaka University Graduate School of Medicine, Osaka, Japan.
authors.forename: Yuzuru
authors.initials: Y
authors.lastname: Kanakura
authors.affiliation: Department of Genetic and Molecular Pathology, Centre for Cancer Biology, SA Pathology, Adelaide, Australia ; School of Pharmacy and Medical Science, University of South Australia, Adelaide, Australia ; School of Medicine, University of Adelaide, Adelaide, Australia ; School of Molecular and Biomedical Science, University of Adelaide, Adelaide, Australia.
authors.forename: Susan
authors.initials: S
authors.lastname: Branford
authors.affiliation: National Hospital Organization Nagoya Medical Center, Nagoya, Japan.
authors.forename: Tomoki
authors.initials: T
authors.lastname: Naoe
keywords: BCR-ABL1; Chronic myeloid leukemia; Conversion factor; International Scale; Real-time quantitative PCR