Jump to ContentJump to Main Navigation
Show Summary Details
More options …

Clinical Chemistry and Laboratory Medicine (CCLM)

Published in Association with the European Federation of Clinical Chemistry and Laboratory Medicine (EFLM)

Editor-in-Chief: Plebani, Mario

Ed. by Gillery, Philippe / Greaves, Ronda / Lackner, Karl J. / Lippi, Giuseppe / Melichar, Bohuslav / Payne, Deborah A. / Schlattmann, Peter

IMPACT FACTOR 2018: 3.638

CiteScore 2018: 2.44

SCImago Journal Rank (SJR) 2018: 1.191
Source Normalized Impact per Paper (SNIP) 2018: 1.205

See all formats and pricing
More options …
Volume 38, Issue 9


Multiplex In-cell Reverse Transcription-Polymerase Chain Reaction for the Simultaneous Detection of p210 and p190 BCR-ABL mRNAs in Chronic Myeloid Leukemia and Philadelphia-Positive Acute Lymphoblastic Leukemia Cell Lines

Soon Pal Suh / Seung Jung Kee / Woo Hyun Lim / Jeong Won Song / Sang Khoo Lee / Jong Phil Kim / Jong Hee Shin / Dong Wook Ryang
Published Online: 2005-06-01 | DOI: https://doi.org/10.1515/CCLM.2000.138


We designed a novel multiplex in-cell reverse transcription- polymerase chain reaction method for the simultaneous detection and differentiation of p190 and p210 BCR-ABL mRNAs within single cells from the human chronic myeloid leukemia and Philadelphia positive acute lymphoblastic leukemia. Human K562 chronic myeloid leukemia and SUP B-15 Ph+ acute lymphoblastic leukemia cell lines were used as positive controls for p210 and p190 BCR-ABL mRNAs, respectively. HL60 cell line was used as a negative control. After the leukemia cells were fixed and permeabilized, without extracting nucleic acids, the mRNAs were reverse transcribed to cDNAs, and the cDNAs were amplified by multiplex polymerase chain reaction with fluorescent primers specific for p190 and p210 BCR-ABL mRNAs. After transfer onto glass slides by cytospin, the amplified cells were detected by fluorescence microscopy. Fluorescence microscopy after propidium iodide or 4′,6-diamidino-2-phenylindone counterstaining showed that the positive K562 cells exhibited a yellow-green fluorescent cytoplasm around a red nucleus, and that the positive SUP B-15 cells exhibited an orange cytoplasm around a blue nucleus. Only the red or blue nucleus was visible in respective negative HL60 cells. The specificity of amplification was confirmed by the absence of a signal when control experiments were performed either with RNase digestion of mRNA or without reverse transcriptase/Taq polymerase. We conclude that the multiplex in-cell reverse transcription-polymerase chain reaction method is capable of simultaneously detecting and differentiating the p210 and p190 BCR-ABL mRNAs of chronic myeloid leukemia and Philadelphia-positive acute lymphoblastic leukemia cells, and that it may be useful in quantitatively monitoring the minimal residual disease during therapy.

About the article

Published Online: 2005-06-01

Published in Print: 2000-09-18

Citation Information: Clinical Chemistry and Laboratory Medicine, Volume 38, Issue 9, Pages 939–944, ISSN (Print) 1434-6621, DOI: https://doi.org/10.1515/CCLM.2000.138.

Export Citation

Citing Articles

Here you can find all Crossref-listed publications in which this article is cited. If you would like to receive automatic email messages as soon as this article is cited in other publications, simply activate the “Citation Alert” on the top of this page.

Manuel Salto-Tellez, Suresh G. Shelat, Bernice Benoit, Hanna Rennert, Martin Carroll, Debra G.B. Leonard, Peter Nowell, and Adam Bagg
The Journal of Molecular Diagnostics, 2003, Volume 5, Number 4, Page 231
Jacques Chasseriau, Jérôme Rivet, Frédéric Bilan, Jean-Claude Chomel, François Guilhot, Nicolas Bourmeyster, and Alain Kitzis
The Journal of Molecular Diagnostics, 2004, Volume 6, Number 4, Page 343

Comments (0)

Please log in or register to comment.
Log in