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An international study to standardize the detection and quantitation of BCR-ABL transcripts from stabilized peripheral blood preparations by quantitative RT-PCR

Lookup NU author(s): Dr Feng Lin, Professor Stephen O'Brien


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Due to the lack of comparability of BCR-ABL mRNA quantification results generated by various methodologies in different laboratories, an international multicenter trial was started with the participation of six laboratories (platforms: LightCycler™, LC, n=3; TaqMan™, TM, n=3). One hundred and eighty-six PB samples derived from healthy donors were spiked with serial dilutions (1:20 to 1:2×106) of b2a2, b3a2 or e1a2 BCR-ABL positive white blood cells (WBC) from leukemic patients. After PAXgene™ stabilization, blinding, freezing and distribution, standardized RNA extraction, cDNA synthesis, PCR protocols and data evaluation were carried out. There was no significant difference in the results achieved using LC and TM technologies, but a considerable overall variation (CV=0.74 for ratios BCR-ABL/ABL). Up to a dilution of 1:1,000, 27/30 of the 2.5 mL samples tested positive. For higher dilutions, a PB volume of 5 or 10 ml was required to improve sensitivity. The study showed the feasibility of RQ-PCR standardization independent of the PCR machine used. ©2007 Ferrata Storti Foundation.

Publication metadata

Author(s): Müller MC, Saglio G, Lin F, Pfeifer H, Press RD, Tubbs RR, Paschka P, Gottardi E, O'Brien SG, Ottmann OG, Stockinger H, Wieczorek L, Merx K, König H, Schwindel U, Hehlmann R, Hochhaus A

Publication type: Article

Publication status: Published

Journal: Haematologica

Year: 2007

Volume: 92

Issue: 7

Pages: 970-973

Print publication date: 01/07/2007

ISSN (print): 0390-6078

ISSN (electronic): 1592-8721


DOI: 10.3324/haematol.11172

PubMed id: 17606448


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