title: Prospective BCR-ABL analysis by polymerase chain reaction (RT-PCR) in adult acute B-lineage lymphoblastic leukemia: reliability of RT-nested-PCR and comparison to cytogenetic data.
abstract: The reliability of routine BCR-ABL RT-nested-PCR was evaluated in 1453 B-lineage ALL or hybrid leukemia at initial diagnosis by RT-nested-PCR. All BCR-ABL-positive (n = 642) and 176 BCR-ABL-negative samples underwent a second RT-PCR. In 518 patients, karyotyping and/or FISH was compared to the BCR-ABL status. The second RT-PCR revealed in 155/642 initially positive samples a divergent result (153 BCR-ABL-negative, two other transcripts) that in most cases turned out to be caused by contaminations in the first RT-nested-PCR. Confirmatory RT-PCR detected 2/176 false negative first RT-nested-PCR results. Thirty-nine specimens remained ambiguous despite different RT-PCR approaches. As far as cytogenetic evaluation and FISH is available (n = 23), the majority but not all patients with an ambiguous RT-PCR result were Ph-negative (n = 18). RT-nested-PCR and cytogenetics yielded in 346 of 383 evaluable samples a concordant result. Differing results are given and account in part to the lower sensitivity of karyotyping. Taken together, confirmed RT-PCR detected BCR-ABL fusion transcripts consistently in 487 out of 1453 ALL samples (c-ALL: 43%, pre-B ALL: 34%, pro-B ALL: 5%, B-ALL: 0%, hybrid leukemia: 5/11). Since false positive initial RT-nested-PCR data were frequent, either confirmatory second RT-PCR or FISH analysis is warranted to guarantee sensitive and reliable results of utmost clinical relevance.
authors.affiliation: Department of Hematology, University Hospital Benjamin Franklin, Berlin, Germany.
authors.forename: B
authors.initials: B
authors.lastname: Gleissner
authors.forename: H
authors.initials: H
authors.lastname: Rieder
authors.forename: E
authors.initials: E
authors.lastname: Thiel
authors.forename: C
authors.initials: C
authors.lastname: Fonatsch
authors.forename: L A
authors.initials: LA
authors.lastname: Janssen
authors.forename: B
authors.initials: B
authors.lastname: Heinze
authors.forename: J W
authors.initials: JW
authors.lastname: Janssen
authors.forename: C
authors.initials: C
authors.lastname: Schoch
authors.forename: N
authors.initials: N
authors.lastname: Goekbuget
authors.forename: J
authors.initials: J
authors.lastname: Maurer
authors.forename: D
authors.initials: D
authors.lastname: Hoelzer
authors.forename: C R
authors.initials: CR
authors.lastname: Bartram
mesh_terms: D000328:Adult; D002051:Burkitt Lymphoma / Q000175:diagnosis* / Q000235:genetics; D020732:Cytogenetic Analysis / Q000592:standards; D003951:Diagnostic Errors; D016044:Fusion Proteins, bcr-abl / Q000032:analysis / Q000235:genetics*; D006801:Humans; D010677:Philadelphia Chromosome; D054198:Precursor Cell Lymphoblastic Leukemia-Lymphoma / Q000175:diagnosis* / Q000235:genetics; D011446:Prospective Studies; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000592:standards*; D012680:Sensitivity and Specificity
chemical_list: D016044:Fusion Proteins, bcr-abl