JMD ASIP MEMBERSHIP
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS

This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Purchase Article
Right arrow View Shopping Cart
Services
Right arrow Similar articles in this journal
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 Google Scholar
Google Scholar
Right arrow Articles by Millson, A.
Right arrow Articles by Lyon, E.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Millson, A.
Right arrow Articles by Lyon, E.
JMD 2003, Vol. 5, No. 3
Copyright © 2003 American Society for Investigative Pathology & Association for Molecular Pathology

Comparison of Two Quantitative Polymerase Chain Reaction Methods for Detecting HER2/neu Amplification

Alison Millson*, Arminda Suli{dagger}, Leah Hartung*, Steve Kunitake{ddagger}, Ann Bennett{ddagger}, Mary C. Lowry Nordberg§, Wedad Hanna, Carl T. Wittwer*{dagger}, Arun Seth and Elaine Lyon*{dagger}

From the Associated Regional and University Pathologists (ARUP) Institute for Clinical and Experimental Pathology, * Salt Lake City, Utah; the Department of Pathology, {dagger} University of Utah School of Medicine, Salt Lake City, Utah; Arcturus, {ddagger} Mountain View, California; the Department of Pathology and Feist-Weiller Cancer Center, § Louisiana State University Health Sciences Center, Shreveport, Louisiana; and Sunnybrook and Women’s College, Health Sciences Center, Toronto, Canada

Two quantitative polymerase chain reaction (PCR) methods for HER2/neu gene quantification were evaluated for implementation into a clinical laboratory. Assays were developed using sequence-specific hybridization probes to detect a target (HER2/neu) and a reference gene (ß-globin) simultaneously. One method utilizes real-time quantification while the second uses internal competitors and melting curves to quantify the unknown sample. These two methods were evaluated using three cell lines and 97 breast tumor samples. Two hundred ninety-four samples were subsequently evaluated using the real-time quantification and immunohistochemical (IHC) staining. Real-time PCR gave HER2/neu gene doses of 10 for SKBR3 and 2 for T47D while the competitive PCR gave doses of 11 for SKBR3 and 2.2 for T47D. Both methods produced coefficients of variation (CV) of less than 3% for within-run and less than 6% for between-run analysis. Examination of 97 breast tumors found a correlation of r = 0.974 between the two methods. IHC and PCR results agreed for 234 of the subsequent 294 samples analyzed (79% concordance). A subset of ten discrepant samples was microdissected. After microdissection all ten were positive by PCR, thus resolving the discrepancy. Real-time quantification and microdissection is useful clinically for HER2/neu quantification. Its ease of use and broad dynamic range allows screening for amplification of HER2/neu.




This article has been cited by other articles:


Home page
 Clin. Cancer Res.Home page
Y. Katsumi, Y. Kuwahara, S. Tamura, K. Kikuchi, O. Otabe, K. Tsuchiya, T. Iehara, H. Kuroda, H. Hosoi, and T. Sugimoto
Trastuzumab Activates Allogeneic or Autologous Antibody-Dependent Cellular Cytotoxicity against Malignant Rhabdoid Tumor Cells and Interleukin-2 Augments the Cytotoxicity
Clin. Cancer Res., February 15, 2008; 14(4): 1192 - 1199.
[Abstract] [Full Text] [PDF]

Home page
 JNCI J Natl Cancer InstHome page
A. McCabe, M. Dolled-Filhart, R. L. Camp, and D. L. Rimm
Automated Quantitative Analysis (AQUA) of In Situ Protein Expression, Antibody Concentration, and Prognosis
J Natl Cancer Inst, December 21, 2005; 97(24): 1808 - 1815.
[Abstract] [Full Text] [PDF]


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Copyright © 2003 by the American Society for Investigative Pathology and the Association for Molecular Pathology.