Quantitation of Plasma Circulating DNA Using Quantitative PCR for the Detection of Hepatocellular Carcinoma

Abstract

Circulating DNA is a potential biomarker for tumor diagnosis and prognosis. This study was aimed to quantify the circulating DNA in plasma from patients with hepatocellular carcinoma (HCC) using quantitative PCR and evaluate its potential clinical value. Blood samples were collected from 72 patients with HCC, 37 with liver cirrhosis or chronic hepatitis and 41 healthy volunteers. Plasma DNA was extracted and quantified by a real-time quantitative PCR method. The diagnostic and prognostic value of plasma DNA analysis for HCC was evaluated. DNA levels in the HCC plasma (median: 173 ng/mL) were significantly higher than those in the healthy controls (9 ng/mL) or control benign patients (46 ng/mL) (P < 0.001). The area under the receiver-operation characteristic (ROC) curve (AUC) assessing plasma DNA was 0.949 for healthy controls and 0.874 for control patients. Plasma DNA detection could discriminate HCC from normal controls with 90.2% sensitivity and 90.3% specificity at the cut-off value of 18.2 ng/mL. Combined ROC analyses using plasma DNA and serum AFP revealed an elevated AUC of 0.974 with 95.1% sensitivity and 94.4% specificity in discriminating HCC from normal controls. The plasma DNA levels were positively associated with tumor size (P = 0.012), and were significantly elevated in HCC patients with intrahepatic spreading or vascular invasion (P = 0.035). The overall survival time of patients with high plasma DNA levels showed a shortened tread when compared with that of patients with low plasma DNA concentrations (P = 0.071). Plasma DNA may be a valuable noninvasive tool for the detecting and predicting the metastasis potential of HCC; and the prognostic value of plasma DNA needed further investigation.

Performance of different biomarkers for the management of hepatocellular carcinoma

“…Moreover, plasma DNA and serum AFP revealed an elevated AUC of 0.974 with 95.1% sensitivity and 94.4% specificity in discriminating HCC from normal controls. Furthermore, the plasma DNA levels were positively associated with tumor size and vascular invasion (P = 0.012 and P = 0.035 respectively) [77] . However further studies are needed due to the controversial results related to the methodology used.…”

“…Recently, several studies have demonstrated that ctDNA could be a non-invasive potential biomarker [54,75] . ctDNA is highly specific and could be detected easier compared to CTCs purification, thus it could be an ideal source for the early diagnosis or as recurrence biomarker [76,77] .…”

“…Several studies have shown that HCC patients have large amounts of cfDNA being these associated with the degree of malignancy (poorer prognosis) and size of the tumor size [77,78] . Huang et al [77] showed that plasma DNA detection was able to discriminate HCC from normal controls with 90.2% sensitivity and 90.3% specificity and AUC was 0.949 (95% CI) (measured by real-time quantitative PCR method).…”

“…Huang et al [77] showed that plasma DNA detection was able to discriminate HCC from normal controls with 90.2% sensitivity and 90.3% specificity and AUC was 0.949 (95% CI) (measured by real-time quantitative PCR method). Moreover, plasma DNA and serum AFP revealed an elevated AUC of 0.974 with 95.1% sensitivity and 94.4% specificity in discriminating HCC from normal controls.…”

Extracellular Nucleic Acids and Cancer

“…progression vs. non-progression) with an almost 80 % sensitivity [148]. When plasma cfmiRNA was profiled from 12 multiple myeloma patients and eight healthy controls using TaqMan low density arrays (TLDAs) in a different set of patients, it was found that high levels of cfmiR-20a and cfmiR-148a were related to shorter relapse-free survival [149]. All these data show some promise but they need to be confirmed in studies including higher patient numbers and longer follow-up times.…”