PMID- 11260080
OWN - NLM
STAT- MEDLINE
DCOM- 20010412
LR  - 20190705
IS  - 0007-1048 (Print)
IS  - 0007-1048 (Linking)
VI  - 112
IP  - 3
DP  - 2001 Mar
TI  - Expression of bcr-abl mRNA in individual chronic myelogenous leukaemia cells as 
      determined by in situ amplification.
PG  - 749-59
AB  - We present the results of a novel method developed for evaluation of in situ 
      amplification, a molecular genetic method at the cellular level. Reverse 
      transcription polymerase chain reaction (RT-PCR) was used to study bcr-abl 
      transcript levels in individual cells from patients with chronic myelogenous 
      leukaemia (CML). After hybridizing a fluorochrome-labelled probe to the 
      cell-bound RT-PCR product, bcr-abl mRNA-positive cells were determined using 
      image analysis. A dilution series of bcr-abl-positive BV173 into normal cells 
      showed a good correlation between expected and actual values. In 25 CML samples, 
      the percentage of in situ PCR-positive cells showed an excellent correlation with 
      cytogenetic results (r = 0.94, P < 0.0001), interphase fluorescence in situ 
      hybridization (FISH) (r = 0.95, P = 0.001) and hypermetaphase FISH (r = 0.81, P < 
      0.001). The fluorescence intensity was higher in residual CML cells after 
      interferon (IFN) treatment than in newly diagnosed patients (P = 0.004), and was 
      highest in late-stage CML resistant to IFN therapy and lowest in CML blast crisis 
      (P = 0.001). Mean fluorescence values correlated with bcr-abl protein levels, as 
      determined by Western blot analysis (r = 0.62). Laser scanning cytometry allowing 
      automated analysis of large numbers of cells confirmed the results. Thus, 
      fluorescence in situ PCR provides a novel and quantitative approach for 
      monitoring tumour load and bcr-abl transcript levels in CML.
FAU - Pachmann, K
AU  - Pachmann K
AD  - The University of Texas M.D. Anderson Cancer Center, Department of Molecular 
      Haematology and Therapy, 1515 Holcombe Boulevard, Houston, TX 77030, USA.
FAU - Zhao, S
AU  - Zhao S
FAU - Schenk, T
AU  - Schenk T
FAU - Kantarjian, H
AU  - Kantarjian H
FAU - El-Naggar, A K
AU  - El-Naggar AK
FAU - Siciliano, M J
AU  - Siciliano MJ
FAU - Guo, J Q
AU  - Guo JQ
FAU - Arlinghaus, R B
AU  - Arlinghaus RB
FAU - Andreeff, M
AU  - Andreeff M
LA  - eng
GR  - CA16672/CA/NCI NIH HHS/United States
GR  - CA4963/CA/NCI NIH HHS/United States
GR  - CA55164/CA/NCI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - England
TA  - Br J Haematol
JT  - British journal of haematology
JID - 0372544
RN  - 0 (RNA, Messenger)
RN  - EC 2.7.10.2 (Fusion Proteins, bcr-abl)
SB  - IM
MH  - Analysis of Variance
MH  - Blotting, Western
MH  - Fusion Proteins, bcr-abl/analysis/*genetics
MH  - Humans
MH  - Image Processing, Computer-Assisted
MH  - In Situ Hybridization, Fluorescence
MH  - Leukemia, Myelogenous, Chronic, BCR-ABL Positive/*genetics/therapy
MH  - Metaphase
MH  - RNA, Messenger/*analysis
MH  - Remission Induction
MH  - Reverse Transcriptase Polymerase Chain Reaction
MH  - Tumor Cells, Cultured
EDAT- 2001/03/22 10:00
MHDA- 2001/04/17 10:01
CRDT- 2001/03/22 10:00
PHST- 2001/03/22 10:00 [pubmed]
PHST- 2001/04/17 10:01 [medline]
PHST- 2001/03/22 10:00 [entrez]
AID - bjh2610 [pii]
AID - 10.1111/j.1365-2141.2001.02510.x [doi]
PST - ppublish
SO  - Br J Haematol. 2001 Mar;112(3):749-59. doi: 10.1111/j.1365-2141.2001.02510.x.