Blood Res 2017; 52(2): 112-118  https://doi.org/10.5045/br.2017.52.2.112
Evaluation of a new flow cytometry based method for detection of BCR-ABL1 fusion protein in chronic myeloid leukemia
Swati Dasgupta1, Ujjal K Ray2, Arpita Ghosh Mitra4, Deboshree M. Bhattacharyya1, Ashis Mukhopadhyay3, Priyabrata Das1, Sudeshna Gangopadhyay1, Sudip Roy4, and Soma Mukhopadhyay1

1Department of Molecular Biology and Hematology, Netaji Subhas Chandra Bose Cancer Research Institute, West Bengal, India.

2Department of Pathology, Netaji Subhas Chandra Bose Cancer Research Institute, West Bengal, India.

3Department of Hemato-Oncology, Netaji Subhas Chandra Bose Cancer Research Institute, West Bengal, India.

4Department of HLA & Molecular Lab, Medica Superspeciality Hospital, West Bengal, India.

Correspondence to: Soma Mukhopadhyay, Ph.D. Department of Molecular Biology and Hematology, Netaji Subhas Chandra Bose Cancer Research Institute, 16 A Park Lane, Kolkata 700016, India. s.amukhopadhyay@gmail.com
Received: September 27, 2016; Revised: November 17, 2016; Accepted: January 6, 2017; Published online: June 22, 2017.
© The Korean Journal of Hematology. All rights reserved.

Abstract

Background

Philadelphia chromosome, a hallmark of chronic myeloid leukemia (CML), plays a key role in disease pathogenesis. It reflects a balanced reciprocal translocation between long arms of chromosomes 9 and 22 involving BCR and ABL1 genes, respectively. An accurate and reliable detection of BCR-ABL fusion gene is necessary for the diagnosis and monitoring of CML. Previously, many technologies, most of which are laborious and time consuming, have been developed to detect BCR-ABL chimeric gene or chromosome.

Methods

A new flow cytometric immunobead assay was used for detection of BCR-ABL fusion proteins and applicability, sensitivity, reliability, efficacy and rapidity of this method was evaluated.

Results

From February 2009 to January 2014, a total 648 CML patients were investigated for the status of BCR-ABL1 protein. Among them, 83 patients were enrolled for comparative study of BCR-ABL1 positivity by three routinely used procedures like karyotyping, and quantitative real time PCR (RT-PCR) as well as immunobead flow cytometry assay. BCR-ABL protein analysis was found consistent, more sensitive (17% greater sensitivity) and reliable than the conventional cytogenetics, as flow cytometry showed 95% concordance rate to RT-PCR.

Conclusion

BCR-ABL fusion protein assay using a new flow cytometric immunobead might be useful in the diagnosis and monitoring CML patients.

Keywords: Chronic myeloid leukemia, BCR-ABL1, Philadelphia chromosome, Flow cytometry


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