Journal of Cancer Research and Clinical Oncology

, Volume 127, Issue 8, pp 489–494

Immunobead RT-PCR versus regular RT-PCR amplification of CEA mRNA in peripheral blood

  • Seungyoon Park
  • Byunghun Lee
  • Ikjung Kim
  • Inhak Choi
  • Kwanhee Hong
  • Yunsick Ryu
  • Junghyo Rhim
  • Juhyun Shin
  • Sang-Chul Park
  • Hongkeun Chung
  • Junho Chung
ORIGINAL PAPER

DOI: 10.1007/s004320100239

Cite this article as:
Park, S., Lee, B., Kim, I. et al. J Cancer Res Clin Oncol (2001) 127: 489. doi:10.1007/s004320100239

Abstract

Purpose: The reverse transcription polymerase chain reaction (RT-PCR) amplification of tumor-specific mRNA has been used for the detection of cancer cells in peripheral blood. More recently, an immunomagnetic isolation and reverse transcription polymerase chain reaction (immunobead RT-PCR) was developed which has reportedly significant advantages over the previous RT-PCR analysis. In our study, we compared these two methods using a model set of peripheral blood containing tumor cells under standardized conditions. Material and methods: In order to compare the false positive rate, normal peripheral blood samples from five volunteers were analyzed by both methods. A model set of peripheral blood containing tumor cells was established by adding SNUC4 human colon cancer cells to peripheral blood collected from normal volunteers not showing any nonspecific bands upon electrophoresis of the PCR products. RT-PCR amplification of carcinoembryonic antigen (CEA) mRNA was done with total RNA and mRNA prepared from this model sample. In immunobead RT-PCR analysis, mRNA was prepared from the cells isolated with anti-CEA antibody-coated magnetic beads or anti-Ber-EP4 antibody-coated magnetic beads before the RT-PCR analysis. Result: The immunobead RT-PCR yielded no non-specific band, while the regular RT-PCR using total RNA did show non-specific band formation in all five samples. When mRNA rather than total RNA was used, nonspecific bands were formed in three of the five samples. Immunobead RT-PCR allowed the detection of 101 tumor cells in 1 ml of peripheral blood. The regular RT-PCR analysis had a detection limit of 102 tumor cells in 1 ml of peripheral blood. Conclusion: The immunobead RT-PCR proved to be more sensitive and specific than the regular RT-PCR at least in our model system.

Keywords CEA RT-PCR Immunobead RT-PCR Colon cancer Immunomagnetic separation 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

Copyright information

© Springer-Verlag Berlin Heidelberg 2001

Authors and Affiliations

  • Seungyoon Park
    • 1
  • Byunghun Lee
    • 1
  • Ikjung Kim
    • 2
  • Inhak Choi
    • 3
  • Kwanhee Hong
    • 4
  • Yunsick Ryu
    • 5
  • Junghyo Rhim
    • 5
  • Juhyun Shin
    • 5
  • Sang-Chul Park
    • 5
  • Hongkeun Chung
    • 5
  • Junho Chung
    • 5
  1. 1.Department of Biochemistry, Dongguk University School of Medicine, 707, Suk-Jang Dong, KyungJu, Kyung-Buk, 780-714, KoreaKR
  2. 2.Department of Microbiology, Dongguk University School of Medicine, 707, Suk-Jang Dong, KyungJu, Kyung-Buk, KoreaKR
  3. 3.Department of Microbiology, Inje University College of Medicine, Pusan 614-735, KoreaKR
  4. 4.Department of Surgery, Inje University College of Medicine, Pusan, KoreaKR
  5. 5.Department of Biochemistry and Molecular Biology and Cancer Research Institute, Seoul National University College of Medicine, Seoul 110-799, Korea Tel.: +82-2-36687440; Fax: +82-2-7444534 e-mail: jjhchung@chollian.netKR

Personalised recommendations