Quantitation of c-erbB-2 in patients with breast cancer by using fluorogenic quantitative polymerase chain reaction method
- VernacularTitle:乳腺癌患者c-erBb-2基因荧光定量PCR方法的研究
- Author:
Donghai LIU
;
Leming ZHANG
;
Shun ZHANG
;
Liangang MAO
;
Weiyun FENG
- Publication Type:Journal Article
- Keywords:
Breast neoplasms;
Genes, c-erbB-2;
Fluorogenic quantitative polymerase chain reaction
- From:
Chinese Journal of Pathophysiology
1986;0(02):-
- CountryChina
- Language:Chinese
-
Abstract:
AIM: To establish a fluorogenic quantitative polymerase chain reaction (FQ-PCR) method for the routine examination of c-erbB-2 gene expression in breast cancer. METHODS: The c-erbB-2 standard gene was obtained by in vitro amplification of cloned c-erbB-2 fragment in plasmid PGEM-T easy vector. FQ-PCR product was detected by using a 7700 ABI PRISM sequence detector system and c-erbB-2 standard curve was obtained to quantity c-erbB-2 in unknown samples. RESULTS: “S” kinetics curve of FQ-PCR amplification was generated by relating the fluorescence signal intensity (△Rn) to the cycle number. The standard curve of c-erbB-2 was constructed by the linear relationship between the cycle threshold (ct) and the log of starting copy number. The high correlation (0.999) revealed the reliability of FQ-PCR. CONCLUSION: The FQ-PCR is a rapid, sensitive, reliable method for quantity of c-erbB-2 gene expression.