Clinical Chemistry
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     

Clinical Chemistry 44: 1161-1169, 1998;
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in Web of Science
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Web of Science (30)
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by O'Connell, C. D.
Right arrow Articles by Hoon, D. S. B.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by O'Connell, C. D.
Right arrow Articles by Hoon, D. S. B.
Related Collections
Right arrow Molecular Diagnostics and Genetics
Right arrow Proteomics and Protein Markers
Right arrow Automation and Analytical Techniques
(Clinical Chemistry. 1998;44:1161-1169.)
© 1998 American Association for Clinical Chemistry, Inc.

Molecular Diagnostics and Genetics

Detection of tyrosinase mRNA in melanoma by reverse transcription-PCR and electrochemiluminescence

Catherine D. O'Connell1,a, Agnes Juhasz1, Christine Kuo2, Dennis J. Reeder1, and Dave S. B. Hoon2

1 Biotechnology Division, Chemical Science and Technology Laboratory, National Institute of Standards and Technology, Gaithersburg, MD 20899.

2 Department of Molecular Oncology, John Wayne Cancer Institute, Santa Monica, CA 90404.
a Author for correspondence. Fax 310-330-3447; e-mail coc{at}nist.gov.

Increased sensitivity and improved quantitation of analytical tests used in biotechnology and clinical chemistry are goals of many laboratories. We have used tyrosinase primers to specifically amplify by RT-PCR the tyrosinase mRNA expressed by the M12 melanoma cell line in a background of mRNA from breast cancer cells. An electrochemiluminescence detection procedure was used as a readout system for this study. Biotinylated post-PCR cDNA samples were hybridized to a tris(2,2'-bipyridine)ruthenium(II) (TBR) chelate-labeled oligonucleotide probe, and the hybrid was subsequently captured by streptavidin-coated Dynabeads®. When either the QPCR System 5000 or the Origen 1 Analyzer System were used, the luminescence emitted by the TBR-chelate of the captured specific post-PCR product was assessed. Tyrosinase-specific mRNA isolated from ~1–10 melanoma cells in a background of 107 cells could be detected. We improved the sensitivity and logistics of the assay through the use of rTth for reverse transcription and amplification. Tyrosinase mRNA was detected in blood from 7 of 16 melanoma patients, whereas none of the 5 healthy donor bloods were positive (P = 0.01; Wilcoxon test).




The following articles in journals at HighWire Press have cited this article:


Home page
 JCOHome page
C. T. Kuo, D. S.B. Hoon, H. Takeuchi, R. Turner, H.-J. Wang, D. L. Morton, and B. Taback
Prediction of Disease Outcome in Melanoma Patients by Molecular Analysis of Paraffin-Embedded Sentinel Lymph Nodes
J. Clin. Oncol., October 1, 2003; 21(19): 3566 - 3572.
[Abstract] [Full Text] [PDF]

Home page
 Clin. Cancer Res.Home page
R. A. Wascher, K. T. Huynh, A. E. Giuliano, N. M. Hansen, F. R. Singer, D. Elashoff, and D. S. B. Hoon
Stanniocalcin-1: A Novel Molecular Blood and Bone Marrow Marker for Human Breast Cancer
Clin. Cancer Res., April 1, 2003; 9(4): 1427 - 1435.
[Abstract] [Full Text] [PDF]

Home page
 Cancer Res.Home page
B. Taback, A. D. Chan, C. T. Kuo, P. J. Bostick, H.-J. Wang, A. E. Giuliano, and D. S. B. Hoon
Detection of Occult Metastatic Breast Cancer Cells in Blood by a Multimolecular Marker Assay: Correlation with Clinical Stage of Disease
Cancer Res., December 1, 2001; 61(24): 8845 - 8850.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Pathol.Home page
D. S. B. Hoon, C. T. Kuo, S. Wen, H. Wang, L. Metelitsa, C. P. Reynolds, and R. C. Seeger
Ganglioside GM2/GD2 Synthetase mRNA Is a Marker for Detection of Infrequent Neuroblastoma Cells in Bone Marrow
Am. J. Pathol., August 1, 2001; 159(2): 493 - 500.
[Abstract] [Full Text]

Home page
 Clin. Chem.Home page
I. Miyashiro, C. Kuo, K. Huynh, A. Iida, D. Morton, A. Bilchik, A. Giuliano, and D. S.B. Hoon
Molecular Strategy for Detecting Metastatic Cancers with Use of Multiple Tumor-specific MAGE-A Genes
Clin. Chem., March 1, 2001; 47(3): 505 - 512.
[Abstract] [Full Text] [PDF]

Home page
 Arch DermatolHome page
H. Tsao, U. Nadiminti, A. J. Sober, and M. Bigby
A Meta-analysis of Reverse Transcriptase-Polymerase Chain Reaction for Tyrosinase mRNA as a Marker for Circulating Tumor Cells in Cutaneous Melanoma
Arch Dermatol, March 1, 2001; 137(3): 325 - 330.
[Abstract] [Full Text] [PDF]

Home page
 Cancer Res.Home page
D. S. B. Hoon, P. Bostick, C. Kuo, T. Okamoto, H.-J. Wang, R. Elashoff, and D. L. Morton
Molecular Markers in Blood as Surrogate Prognostic Indicators of Melanoma Recurrence
Cancer Res., April 1, 2000; 60(8): 2253 - 2257.
[Abstract] [Full Text]

Home page
 Clin. Chem.Home page
F. Fini, G. Gallinella, S. Girotti, M. Zerbini, and M. Musiani
Development of a Chemiluminescence Competitive PCR for the Detection and Quantification of Parvovirus B19 DNA Using a Microplate Luminometer
Clin. Chem., September 1, 1999; 45(9): 1391 - 1396.
[Abstract] [Full Text] [PDF]


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Copyright © 1998 by the American Association for Clinical Chemistry.