总血清DNA和DNA完整性：乙型肝炎病毒相关肝细胞癌患者的诊

StephenW

Pathology. 2012 Jun;44(4):318-324. doi: 10.1097/PAT.0b013e328353a24c.
Total serum DNA and DNA integrity: diagnostic value in patients with hepatitis B virus-related hepatocellular carcinoma.Chen H1, Sun LY2, Zheng HQ2, Zhang QF3, Jin XM4.
Author information

• 1Department of Urinary Surgery, The Affiliated Tumor Hospital, Harbin Medical University, Harbin, China.
• 2Department of General Surgery, Fourth Affiliated Hospital, Harbin Medical University, Harbin, China.
• 3Department of General Surgery, Fourth Affiliated Hospital, Harbin Medical University, Harbin, China. Electronic address: [email protected].
• 4Department of Pathology, Harbin Medical University, Harbin, China. Electronic address: [email protected].
  

AbstractAIMS: This study aimed to test the diagnostic utility of the total serum cell-free DNA (cfDNA) and DNA integrity index for detection of hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC).
METHODS: We initially evaluated the sodium iodide (NaI) method, Triton/Heat/Phenol (THP) protocol and QIAamp Kit for cfDNA extraction. Then cfDNA was isolated from the sera of 80 patients with HBV-related HCC, 80 patients with chronic HBV infection and 50 healthy subjects, and quantified by realtime quantitative polymerase chain reaction (qPCR) amplification of beta-actin genomic DNA fragments using two sets of primers of 100 and 400 bp. DNA integrity was calculated as the ratio of 400 bp to 100 bp β-actin fragments.
RESULTS: The THP approach was not only superior to the other two methods in terms of DNA quantity, but also was simpler, more rapid, and less costly. Serum DNA integrity in HCC patients was significantly higher than that in HBV patients or healthy controls. As for total cfDNA levels, although a significant difference was found between HCC patients and healthy individuals, no significant difference was found between HBV patients with and without HCC. DNA integrity was associated with tumour size, TNM stage, lymph node and distant metastasis. DNA integrity had a higher sensitivity and specificity in discriminating HCC from HBV patients than total DNA.
CONCLUSIONS: The THP method is preferred for extraction of cfDNA. DNA integrity is a promising molecular biomarker for detecting HCC in patients with chronic HBV infection; it reflects the progression and metastatic potential of the tumour.

Copyright © 2012 Royal College of Pathologists of Australasia. Published by Elsevier B.V. All rights reserved.

KEYWORDS: Circulating DNA; DNA integrity; HCC; real-time PCR; serum

PMID:28193336DOI:10.1097/PAT.0b013e328353a24c

StephenW

病理。 2012 Jun; 44（4）：318-324。 doi：10.1097 / PAT.0b013e328353a24c。
总血清DNA和DNA完整性：乙型肝炎病毒相关肝细胞癌患者的诊断价值。
Chen H1，Sun LY2，Zheng HQ2，Zhang QF3，Jin XM4。
作者信息

    1哈尔滨医科大学附属肿瘤医院泌尿外科，哈尔滨。
    2哈尔滨医科大学第四附属医院普通外科，哈尔滨。
    哈尔滨医科大学第四附属医院普外外科，哈尔滨。电子地址：[email protected]。
    哈尔滨医科大学病理学教研室，哈尔滨。电子地址：[email protected]。

抽象
AIMS：

本研究旨在测试总血清无细胞DNA（cfDNA）和DNA完整性指数的诊断效用，用于检测乙型肝炎病毒（HBV）相关的肝细胞癌（HCC）。
方法：

我们最初评估碘化钠（NaI）方法，Triton /热/苯酚（THP）方案和用于cfDNA提取的QIAamp试剂盒。然后从80名HBV相关HCC患者，80名慢性HBV感染患者和50名健康受试者的血清中分离cfDNA，并通过实时定量聚合酶链反应（qPCR）扩增β-肌动蛋白基因组DNA片段，使用两组引物100和400 bp。 DNA完整性计算为400bp至100bpβ-肌动蛋白片段的比率。
结果：

THP方法不仅在DNA量方面优于其他两种方法，而且更简单，更快速，并且成本更低。 HCC患者的血清DNA完整性显着高于HBV患者或健康对照。至于总cfDNA水平，尽管在HCC患者和健康个体之间发现显着差异，但在具有和不具有HCC的HBV患者之间没有发现显着差异。 DNA完整性与肿瘤大小，TNM期，淋巴结和远处转移相关。 DNA完整性在区分HBV患者的HCC方面具有比总DNA更高的灵敏度和特异性。
结论：

THP方法优选用于提取cfDNA。 DNA完整性是一种有希望的分子生物标志物，用于检测慢性HBV感染患者的HCC;它反映了肿瘤的进展和转移潜力。

版权所有©2012澳大利亚皇家病理学家学院。发表者Elsevier B.V.保留所有权利。
关键词：

循环DNA; DNA完整性; HCC;实时PCR;血清

PMID：
    28193336
DOI：
    10.1097 / PAT.0b013e328353a24c