[癌症患者血清中循环DNA的定量分析]

[Quantitative analysis of circulating DNA in serum of cancer patients].

作者信息

Tu Hong, Gao Hai-feng, Fu Shi-long, Chen Hao

机构信息

Shanghai Cancer Institute/Cancer Institute of Shanghai Jiao-Tong University, Shanghai 200032, China.

出版信息

Zhonghua Zhong Liu Za Zhi. 2004 Oct;26(10):606-8.

PMID:15634521

Abstract

OBJECTIVE

To develop a method for ng quantitation of circulating DNA in serum and explore the value in the diagnosis of cancer.

METHODS

Serum DNA was extracted by commercial "genomic DNA extraction kit" and detected by fluorescent dye (SYBR green I) staining. Loss of heterozygosity (LOH) at BRCA1 (D17S579, D17S855) and p53 (TP53, D17S786) in serum DNA was analyzed by PCR-based method.

RESULTS

SYBR green I dot staining could detect DNA as low as 2 ng. Using this method, we detected serum samples from 483 patients with various types of cancer and 150 healthy individuals. The mean DNA concentration in the normal controls was 22.2 +/- 13.4 ng/ml, while that in cancer patients was 81.3 +/- 98.3 ng/ml (P < 0.001). In 33 ovarian cancer patients with increased DNA level, 27(81.8%) displayed LOH in at least one of the four loci analyzed.

CONCLUSION

Circulating DNA in serum may become additional tumor marker for the diagnosis of cancer.

摘要

目的

建立一种血清中循环DNA的纳克级定量方法，并探讨其在癌症诊断中的价值。

方法

采用市售“基因组DNA提取试剂盒”提取血清DNA，并用荧光染料（SYBR green I）染色进行检测。采用基于聚合酶链反应（PCR）的方法分析血清DNA中乳腺癌1号基因（BRCA1，位点D17S579、D17S855）和p53基因（TP53，位点D17S786）的杂合性缺失（LOH）情况。

结果

SYBR green I斑点染色可检测低至2纳克的DNA。运用该方法，我们检测了483例各类癌症患者及150名健康个体的血清样本。正常对照组的平均DNA浓度为22.2±13.4纳克/毫升，而癌症患者组为81.3±98.3纳克/毫升（P<0.001）。在33例DNA水平升高的卵巢癌患者中，27例（81.8%）在分析的四个位点中至少有一个位点出现杂合性缺失。