PMID- 10462577
OWN - NLM
STAT- Publisher
LR  - 20191120
IS  - 1084-8592 (Print)
IS  - 1084-8592 (Linking)
VI  - 1
IP  - 4
DP  - 1996 Dec
TI  - Detection of BCRabl in Acute Leukemia by Molecular and Cytogenetic Methods.
PG  - 305-313
AB  - Background: The Philadelphia chromosome (Ph), t(9;22)(q34;q11), is detected by 
      karyotyping in a minority of patients with acute leukemia. Ph results in fusion 
      of the c-abl oncogene on chromosome 9 with the breakpoint cluster region BCR gene 
      on chromosome 22. The purpose of this study was to compare reverse 
      trascriptase-polymerase chain reaction (RT-PCR) for BCR/abl fusion to cytogenetic 
      methods for Ph detection in patients with acute leukemia. Methods and Results: 
      Peripheral blood and bone marrow samples from cases of adult acute myelogenous 
      leukemia (AML) and acute lymphoblastic leukemia (ALL) were examined for Ph by 
      RT-PCR, karyotyping, and fluorescence in situ hybridization (FISH). Using total 
      cellular RNA and a single primer pair, cDNA was transcribed, amplified, 
      electorphoresed, and probed for BCR/abl fusion. Patient cells and SUPB15 and K562 
      cell lines were used as breakpoint controls. Karyotyping was done by standard 
      Giemsa banding. FISH was performed on bone marrow smears using digxigenin-labeled 
      DNA probes for major and minor bcr breakpoints (corresponding to involvement of 
      major bcr exons 2/3 and minor bcr exon 1, respectively) and biotin-labeled DNA 
      probes for abl. Rhodamine-conjugated antidigoxigenin and fluorescein-conjugated 
      avidin yielded red and green fluorescent signals, respectively. A total of 32 
      samples from patients with AML were studied, 20 from patients with de novo or 
      relapsed AML and 12 from patients in remission. Five of 32 cases of AML (16%) 
      were RT-PCR+/Ph+ all with major bcr breakpoints between exons 3 and 4. One of the 
      32 cases (3%) was RT-PCR+/Ph+; this case was the only positive remission sample. 
      Of the four T-PCR+/Ph- cases, one showed t(2;17), one showed t(9;11), and two had 
      a normal karyotype. FISH was done in three RT-PCR+ cases, yielding positive 
      results with the major probe in two. A total of 22 samples from patients with ALL 
      were studies, 15 from patients with de novo or relapsed ALL and seven from 
      patient remission. Seven of 22 cases of ALL (32%) were RT-PCR+, four with 
      major-bcr breakpoints between exons 3 and 4, and three with breakpoints in the 
      minor-bcr. Two of the 22 (9%) cases were RT-PCR+/Ph+. Of the five RT-PCR+/Ph- 
      cases, two showed a 22q- but lacked the typical Ph break on chromosome 9, 1 
      showed a 12p-, and two had a normal karyotype. FISH was performed in four RT-PCR+ 
      cases, yielding positive results with the major probe in two cases and with the 
      minor probe in two cases. Conclusions: RT-PCR is more sensitive than karyotyping, 
      detecting masked Ph or translocations not found by cytogenetic analysis. FISH is 
      a helpful adjunctive test when used to confirm BCR/abl fusion in RT-PCR+/Ph- 
      cases.
FAU - Wells, SJ
AU  - Wells SJ
AD  - Department of Pathology and Laboratory Medicine, Emory University School of 
      Medicine, Atlanta, Georgia, USA
FAU - Phillips, CN
AU  - Phillips CN
FAU - Farhi, DC
AU  - Farhi DC
LA  - eng
PT  - Journal Article
PL  - United States
TA  - Mol Diagn
JT  - Molecular diagnosis : a journal devoted to the understanding of human disease 
      through the clinical application of molecular biology
JID - 9614965
EDAT- 1996/12/01 00:00
MHDA- 1999/08/27 00:00
CRDT- 1996/12/01 00:00
PHST- 1996/12/01 00:00 [pubmed]
PHST- 1999/08/27 00:00 [medline]
PHST- 1996/12/01 00:00 [entrez]
AID - S1084859296000240 [pii]
AID - 10.1054/MODI00100305 [doi]
PST - ppublish
SO  - Mol Diagn. 1996 Dec;1(4):305-313. doi: 10.1054/MODI00100305.