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 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.
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 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.
Authors: C J Monjure; C D Tatum; A T Panganiban; M Arainga; V Traina-Dorge; P A Marx; E S Didier Journal: J Med Primatol Date: 2013-11-22 Impact factor: 0.667
Authors: N C P Cross; H E White; D Colomer; H Ehrencrona; L Foroni; E Gottardi; T Lange; T Lion; K Machova Polakova; S Dulucq; G Martinelli; E Oppliger Leibundgut; N Pallisgaard; G Barbany; T Sacha; R Talmaci; B Izzo; G Saglio; F Pane; M C Müller; A Hochhaus Journal: Leukemia Date: 2015-02-05 Impact factor: 11.528
Authors: Anoop Enjeti; Neil Granter; Asma Ashraf; Linda Fletcher; Susan Branford; Philip Rowlings; Susan Dooley Journal: Pathology Date: 2015-10 Impact factor: 5.306
Authors: H White; L Deprez; P Corbisier; V Hall; F Lin; S Mazoua; S Trapmann; A Aggerholm; H Andrikovics; S Akiki; G Barbany; N Boeckx; A Bench; M Catherwood; J-M Cayuela; S Chudleigh; T Clench; D Colomer; F Daraio; S Dulucq; J Farrugia; L Fletcher; L Foroni; R Ganderton; G Gerrard; E Gineikienė; S Hayette; H El Housni; B Izzo; M Jansson; P Johnels; T Jurcek; V Kairisto; A Kizilors; D-W Kim; T Lange; T Lion; K M Polakova; G Martinelli; S McCarron; P A Merle; B Milner; G Mitterbauer-Hohendanner; M Nagar; G Nickless; J Nomdedéu; D A Nymoen; E O Leibundgut; U Ozbek; T Pajič; H Pfeifer; C Preudhomme; K Raudsepp; G Romeo; T Sacha; R Talmaci; T Touloumenidou; V H J Van der Velden; P Waits; L Wang; E Wilkinson; G Wilson; D Wren; R Zadro; J Ziermann; K Zoi; M C Müller; A Hochhaus; H Schimmel; N C P Cross; H Emons Journal: Leukemia Date: 2014-07-18 Impact factor: 11.528
Authors: Alison S Devonshire; Rebecca Sanders; Alexandra S Whale; Gavin J Nixon; Simon Cowen; Stephen L R Ellison; Helen Parkes; P Scott Pine; Marc Salit; Jennifer McDaniel; Sarah Munro; Steve Lund; Satoko Matsukura; Yuji Sekiguchi; Mamoru Kawaharasaki; José Mauro Granjeiro; Priscila Falagan-Lotsch; Antonio Marcos Saraiva; Paulo Couto; Inchul Yang; Hyerim Kwon; Sang-Ryoul Park; Tina Demšar; Jana Žel; Andrej Blejec; Mojca Milavec; Lianhua Dong; Ling Zhang; Zhiwei Sui; Jing Wang; Duangkamol Viroonudomphol; Chaiwat Prawettongsopon; Lina Partis; Anna Baoutina; Kerry Emslie; Akiko Takatsu; Sema Akyurek; Muslum Akgoz; Maxim Vonsky; L A Konopelko; Edna Matus Cundapi; Melina Pérez Urquiza; Jim F Huggett; Carole A Foy Journal: Biomol Detect Quantif Date: 2016-06-06