title: Quantification of residual disease in Philadelphia-positive acute lymphoblastic leukemia: comparison of blood and bone marrow.
abstract: We designed a new semi-quantitative competitor-based PCR assay to assess the amount of p190 BCR-ABL mRNA in patients with Ph-positive ALL. Transcript numbers were compared in 29 paired specimens of blood and marrow collected contemporaneously from 18 patients at differing stages of disease. In general, the numbers of BCR-ABL transcripts detected in marrow in blood were not significantly different (p = 0.1). However, in four samples BCR-ABL transcripts (< 10-1000/micrograms RNA) were detected in the marrow while the blood was negative; the reverse, positive blood and negative marrow, was not seen. In a further three samples the number of BCR-ABL transcripts was more than 10-fold higher in the marrow. We measured the number of ABL transcripts/micrograms RNA in all samples as an internal standard in order to control for variations in sample quality and other parameters. For two out of the four discordant samples in which blood was PCR negative, the number of ABL transcripts/micrograms RNA detected in the marrow was substantially higher than in the blood, suggesting poor quality blood specimens. However, the ratio of BCR-ABL to ABL in marrow and blood was similar for the three discordant samples in which both tissues were PCR positive. We conclude that in general, blood and marrow contain similar BCR-ABL transcript numbers in Ph-positive ALL but some samples are discordant. Marrow is therefore the preferred tissue for residual disease studies. Quantification of ABL mRNA as an internal control is useful in the interpretation of competitive PCR data and may serve as a robust way to standardize results between laboratories.
authors.affiliation: Department of Haematology, Royal Postgraduate Medical School, London, UK.
authors.forename: F
authors.initials: F
authors.lastname: van Rhee
authors.forename: D I
authors.initials: DI
authors.lastname: Marks
authors.forename: F
authors.initials: F
authors.lastname: Lin
authors.forename: R M
authors.initials: RM
authors.lastname: Szydlo
authors.forename: A
authors.initials: A
authors.lastname: Hochhaus
authors.forename: J
authors.initials: J
authors.lastname: Treleaven
authors.forename: C
authors.initials: C
authors.lastname: Delord
authors.forename: N C
authors.initials: NC
authors.lastname: Cross
authors.forename: J M
authors.initials: JM
authors.lastname: Goldman
mesh_terms: D001483:Base Sequence; D001773:Blood Cells / Q000473:pathology*; D001853:Bone Marrow / Q000473:pathology*; D016044:Fusion Proteins, bcr-abl / Q000032:analysis / Q000235:genetics*; D006801:Humans; D008969:Molecular Sequence Data; D018365:Neoplasm, Residual; D014411:Neoplastic Stem Cells / Q000473:pathology*; D010677:Philadelphia Chromosome; D016133:Polymerase Chain Reaction; D054198:Precursor Cell Lymphoblastic Leukemia-Lymphoma / Q000097:blood / Q000235:genetics / Q000473:pathology*; D016315:Proto-Oncogene Proteins c-abl / Q000032:analysis; D012333:RNA, Messenger / Q000032:analysis; D012334:RNA, Neoplasm / Q000032:analysis; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D012333:RNA, Messenger; D012334:RNA, Neoplasm; D016044:Fusion Proteins, bcr-abl; D016315:Proto-Oncogene Proteins c-abl