Quantitative analysis of circulating tumor DNA as a diagnostic marker for breast cancer and its role
10.3760/cma.j.issn.0254-9026.2010.01.012
- VernacularTitle:循环肿瘤DNA定量方法的建立及其临床应用的初步研究
- Author:
Jinghua LI
;
Bo XU
;
Hongyan LI
;
Jiang LI
;
Mei XU
;
Fuyun GAO
;
Qing FANG
- Publication Type:Journal Article
- Keywords:
DNA;
neoplasms;
Breast neoplasms
- From:
Chinese Journal of Geriatrics
2010;29(1):38-41
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish the methods for quantitating the circulating tumor DNA with PicoGreen fluorescent nucleic acid stain and investigate the role of the quantitative analysis in diagnosis of breast cancer. Methods Circulating tumor DNA was isolated from serum with QIAmp blood kit and was quantitated by spectrofluorometry with PicoGreen fluorescent stain. Receiver operator characteristic ( ROC) curve and area under the curve were used to estimate the role of DNA quantification in diagnosis of breast cancer. Results The efficiency of QIAamp blood kit isolating DNA from serum was 37. 8%-46. 2%, average 43. 4%. Circulating tumor DNA concentration as low as 1 ng could be detected by PicoGreen spectrofluorometry. and the detected range was 1-500 ng/0. 2 ml. The median concentration of serum DNA in breast cancer group was (169. 70+ 124. 10) μg/L, and that of healthy control and breast benign group were (54. 30±36. 84) μg/L and (51. 70±29. 04) μg/L, respectively (P<0. 01). The area under the ROC curve was 0. 899 (95% CI: 0.848-0.951), and the sensitivity was 78. 2%, the specificity was 90% by using the cutoff value of 96. 0 ng/ml. Conclusions The concentration of circulating tumor DNA can be efficiently quantitated by PicoGreen spectrofluorometry, which indicates the potential of clinical applicability in breast cancer diagnosis.
