PMID- 15276916
OWN - NLM
STAT- MEDLINE
DCOM- 20050106
LR  - 20191210
IS  - 0036-5513 (Print)
IS  - 0036-5513 (Linking)
VI  - 64
IP  - 5
DP  - 2004
TI  - Quantitative PCR--new diagnostic tool for quantifying specific mRNA and DNA 
      molecules: HER2/neu DNA quantification with LightCycler real-time PCR in 
      comparison with immunohistochemistry and fluorescence in situ hybridization.
PG  - 511-22
AB  - Previously, polymerase chain reaction (PCR) technology has been hampered by its 
      inability to generate quantitative results, a drawback inherent to the high 
      degree of amplification taking place in the reaction. Recently, PCR techniques 
      have been described with the potential of quantifying the amount of mRNA or DNA 
      in biological samples. In this study quantitative PCR was used to investigate the 
      role of the EGF (epidermal growth factor) system in cancer both for measurements 
      of mRNA concentrations and for measurements of the number of copies of specific 
      genes. It is shown that the mRNA expression of a subset of ligands from the EGF 
      system is increased in bladder cancer. Furthermore, measurement of the mRNA 
      concentration gives important information such as the expression of these ligands 
      correlated to the survival of the patients. In addition to the alterations at the 
      mRNA level, changes also can occur at the DNA level in the EGF system. Thus, it 
      has been demonstrated that the number of genes coding for the human epidermal 
      growth factor receptor 2 (HER2) is increased in a number of breast tumors. It is 
      now possible to treat breast cancer patients with a humanized antibody reacting 
      with HER2, and the treatment is considered to be justified if the tumor displays 
      an increased amount of HER2. For this reason there is a need for techniques 
      suitable for HER2 measurements. A LightCycler real-time PCR method used for 
      HER2/neu DNA quantification was evaluated and the results compared with those 
      obtained by immunohistochemistry (IHC) and fluorescence in situ hybridization 
      (FISH). Tumor biopsies were collected from 112 patients diagnosed with early 
      breast cancer from January 1990 to March 1994. The samples were analyzed for HER2 
      DNA amplification by real-time PCR on LightCycler and by FISH and for HER2 
      protein expression by IHC. Inter-assay variation for HER2 measured by LightCycler 
      was 10% (x =3.1; n=17). Amplification > or = 2 was observed in 19% of the 
      patients. Concordance rates between real-time PCR and the other methods were 91% 
      (IHC) and 92% (FISH). The correlation between real-time PCR and FISH was highly 
      significant (p < 0.001). The "LightCycler-HER2/neu DNA quantification kit" 
      produces results with a high level of reproducibility and its ease of use allows 
      rapid screening for amplification of HER2. In this paper useful information is 
      given on how real-time PCR compares with FISH and IHC. The data show that results 
      obtained for amplification of HER2 by real-time PCR on the LightCycler instrument 
      are comparable to results obtained by IHC and FISH.
FAU - Schlemmer, B O
AU  - Schlemmer BO
AD  - Department of Clinical Biochemistry, Aarhus University Hospital, Denmark. 
      boess@akh.aaa.dk
FAU - Sorensen, B S
AU  - Sorensen BS
FAU - Overgaard, J
AU  - Overgaard J
FAU - Olsen, K E
AU  - Olsen KE
FAU - Gjerdrum, L M
AU  - Gjerdrum LM
FAU - Nexo, E
AU  - Nexo E
LA  - eng
PT  - Comparative Study
PT  - Evaluation Study
PT  - Historical Article
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - Scand J Clin Lab Invest
JT  - Scandinavian journal of clinical and laboratory investigation
JID - 0404375
RN  - 0 (DNA, Neoplasm)
RN  - 0 (RNA, Messenger)
SB  - IM
MH  - Awards and Prizes
MH  - Breast Neoplasms/genetics/metabolism/pathology
MH  - Chemistry, Clinical/history
MH  - Clinical Medicine/history
MH  - DNA, Neoplasm/*analysis
MH  - *Genes, erbB-2
MH  - History, 21st Century
MH  - Immunohistochemistry/*methods
MH  - In Situ Hybridization, Fluorescence/*methods
MH  - Light
MH  - RNA, Messenger/*analysis
MH  - Reproducibility of Results
MH  - *Reverse Transcriptase Polymerase Chain Reaction
MH  - Scandinavian and Nordic Countries
MH  - Societies, Medical
EDAT- 2004/07/28 05:00
MHDA- 2005/01/07 09:00
CRDT- 2004/07/28 05:00
PHST- 2004/07/28 05:00 [pubmed]
PHST- 2005/01/07 09:00 [medline]
PHST- 2004/07/28 05:00 [entrez]
AID - 7B2N2P4WW1GAD7U4 [pii]
AID - 10.1080/00365510410002922 [doi]
PST - ppublish
SO  - Scand J Clin Lab Invest. 2004;64(5):511-22. doi: 10.1080/00365510410002922.