Programme for Harmonization to the International Scale in Latin America for BCR-ABL1 quantification in CML patients: findings and recommendations

María Sol Ruiz 1 , María Belén Sánchez 1 , Yuly Masiel Vera Contreras 1 , Evangelina Agrielo 2 , Marta Alonso 3 , María Eugenia Altuna 4 , María Sol Anchordoqui 5 , Mariana Asinari 6 , María Elisa Bonetto 7 , Mauricio Camargo 8 , Isabel Giere 9 , Javier González 10 , Ana Cecilia Granda Alacote 11 , Javier Guerra 12 , Marina Gutiérrez 13 , Cecilia Maldonado 14 , Ricardo Makiya 15 , Gonzalo Manrique 16 , María Eugenia Monaco 17 , Juan Carlos Rozo 18 , Carlos Santamaría 19 , Analía Seravalle 20 , Olga Zea 8 , María Noel Zubillaga 16 , José Mordoh 1 , Irene Larripa 21 ,  and Michele Bianchini 1 , 5 , 21
  • 1 CIO-FUCA, Centro de Investigaciones Oncológicas – Fundación Cáncer, Ciudad Autónoma de Buenos Aires, Buenos Aires, Argentina
  • 2 LEB, Laboratorio de Especialidades Bioquímicas, Bahia Blanca, Buenos Aires, Argentina
  • 3 Hospital Nac. A. Posadas, El Palomar, Buenos Aires, Argentina
  • 4 Clinica Dr. Roberto Raña, Neuquen, Argentina
  • 5 Argenomics, Pilar, Buenos Aires, Argentina
  • 6 Hospital Privado Universitario, Cordoba, Argentina
  • 7 Hospital Dr. G. Rawson, San Juan, Argentina
  • 8 Genética Lab, Medellín, Antioquia, Colombia
  • 9 Fundaleu, Ciudad Autónoma de Buenos Aires, Buenos Aires, Argentina
  • 10 Meyer Lab, Asunción, Paraguay
  • 11 Laboratorios Medicos A Yuen y G Rios, Lima, Peru
  • 12 Nanopharmacia Diagnostica, Ciudad de Mexico, Mexico
  • 13 Stamboulian, Ciudad Autónoma de Buenos Aires, Buenos Aires, Argentina
  • 14 ManLab, Ciudad Autónoma de Buenos Aires, Buenos Aires, Argentina
  • 15 FIBIO, Ciudad Autónoma de Buenos Aires, Buenos Aires, Argentina
  • 16 ASESP, Asociación Española, Montevideo, Uruguay
  • 17 Laboratorio Tucuman, San Miguel de Tucumán, Tucumán, Argentina
  • 18 UDHO, Unidad de Diagnóstico Hemato Oncológico, Cali, Valle del Cauca, Colombia
  • 19 Hospital de Niños Carlos Sáenz Herrera, San José, Costa Rica
  • 20 CIBIC, Rosario, Santa Fe, Argentina
  • 21 IMEX, Instituto de Medicina Experimental, CONICET – Academia Nacional de Medicina, Ciudad Autónoma de Buenos Aires, Buenos Aires, Argentina
María Sol Ruiz
  • CIO-FUCA, Centro de Investigaciones Oncológicas – Fundación Cáncer, Ciudad Autónoma de Buenos Aires, Buenos Aires, Argentina
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, María Belén Sánchez
  • CIO-FUCA, Centro de Investigaciones Oncológicas – Fundación Cáncer, Ciudad Autónoma de Buenos Aires, Buenos Aires, Argentina
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, Yuly Masiel Vera Contreras
  • CIO-FUCA, Centro de Investigaciones Oncológicas – Fundación Cáncer, Ciudad Autónoma de Buenos Aires, Buenos Aires, Argentina
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, Evangelina Agrielo
  • LEB, Laboratorio de Especialidades Bioquímicas, Bahia Blanca, Buenos Aires, Argentina
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, Marta Alonso, María Eugenia Altuna, María Sol Anchordoqui, Mariana Asinari, María Elisa Bonetto, Mauricio Camargo, Isabel Giere, Javier González, Ana Cecilia Granda Alacote, Javier Guerra, Marina Gutiérrez, Cecilia Maldonado, Ricardo Makiya, Gonzalo Manrique, María Eugenia Monaco, Juan Carlos Rozo, Carlos Santamaría, Analía Seravalle, Olga Zea, María Noel Zubillaga, José Mordoh
  • CIO-FUCA, Centro de Investigaciones Oncológicas – Fundación Cáncer, Ciudad Autónoma de Buenos Aires, Buenos Aires, Argentina
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, Irene Larripa
  • IMEX, Instituto de Medicina Experimental, CONICET – Academia Nacional de Medicina, Ciudad Autónoma de Buenos Aires, Buenos Aires, Argentina
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and Michele Bianchini
  • Corresponding author
  • CIO-FUCA, Centro de Investigaciones Oncológicas – Fundación Cáncer, Ciudad Autónoma de Buenos Aires, Buenos Aires, Argentina
  • Argenomics, Pilar, Buenos Aires, Argentina
  • IMEX, Instituto de Medicina Experimental, CONICET – Academia Nacional de Medicina, Ciudad Autónoma de Buenos Aires, Buenos Aires, Argentina
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Abstract

Objectives

The quantitation of BCR-ABL1 mRNA is mandatory for chronic myeloid leukemia (CML) patients, and RT-qPCR is the most extensively used method in testing laboratories worldwide. Nevertheless, substantial variation in RT-qPCR results makes inter-laboratory comparability hard. To facilitate inter-laboratory comparative assessment, an international scale (IS) for BCR-ABL1 was proposed.

Methods

The laboratory-specific conversion factor (CF) to the IS can be derived from the World Health Organization (WHO) genetic reference panel; however, this material is limited to the manufacturers to produce and calibrate secondary reference reagents. Therefore, we developed secondary reference calibrators, as lyophilized cellular material, aligned to the IS. Our purpose was both to re-evaluate the CF in 18 previously harmonized laboratories and to propagate the IS to new laboratories.

Results

Our field trial including 30 laboratories across Latin America showed that, after correction of raw BCR-ABL1/ABL1 ratios using CF, the relative mean bias was significantly reduced. We also performed a follow-up of participating laboratories by annually revalidating the process; our results support the need for continuous revalidation of CFs. All participating laboratories also received a calibrator to determine the limit of quantification (LOQ); 90% of them could reproducibly detect BCR-ABL1, indicating that these laboratories can report a consistent deep molecular response. In addition, aiming to investigate the variability of BCR-ABL1 measurements across different RNA inputs, we calculated PCR efficiency for each individual assay by using different amounts of RNA.

Conclusions

In conclusion, for the first time in Latin America, we have successfully organized a harmonization platform for BCR-ABL1 measurement that could be of immediate clinical benefit for monitoring the molecular response of patients in low-resource regions.

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Clinical Chemistry and Laboratory Medicine ( CCLM) publishes articles on novel teaching and training methods applicable to laboratory medicine. CCLM welcomes contributions on the progress in fundamental and applied research and cutting-edge clinical laboratory medicine. It is one of the leading journals in the field, with an impact factor of over three. CCLM is the official journal of nine national clinical societies and associated with EFLM.

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