Next generation digital PCR measurement of hepatitis B virus copy number in formalin-fixed paraffin-embedded hepatocellular carcinoma tissue.

Abstract

BACKGROUND:

Hepatocellular carcinoma (HCC) is strongly associated with hepatitis B virus (HBV) infection. False-negative results are common in routine serological tests and quantitative real-time PCR because of HBV surface antigen (HBsAg) variation and low HBV copy number. Droplet digital PCR (ddPCR), a next generation digital PCR, is a novel, sensitive, and specific platform that can be used to improve HBV detection.

METHODS:

A total of 131 HCC cases with different tumor stages and clinical features were initially classified with a serological test as HBsAg positive (n = 107) or negative (n = 24) for HBV infection. Next, DNA templates were prepared from the corresponding formalin-fixed paraffin-embedded (FFPE) tissues to determine HBV copy number by ddPCR.

RESULTS:

HBV copy numbers, successfully determined for all clinical FFPE tissues (n = 131), ranged from 1.1 to 175.5 copies/μL according to ddPCR. The copy numbers of HBV were positively correlated with tumor-nodes-metastasis (P = 0.008) and Barcelona-Clinic Liver Cancer (P = 0.045) classification. Moreover, serum cholinesterase correlated with hepatitis B viral load (P = 0.006).

CONCLUSIONS:

HBV infection is a key factor that influences tumorigenesis in HCC by regulating tumor occurrence and development. ddPCR improves the analytical sensitivity and specificity of measurements in nucleic acids at a single-molecule level and is suitable for HBV detection.

• Authors: Jing-Tao Huang, Ying-Juan Liu, Jin Wang, Zhi-Gao Xu, Ying Yang, Fan Shen, Xing-Hui Liu, Xin Zhou, Song-Mei Liu
• Journal: Clinical chemistry (Clin Chem) Vol. 61 Issue 1 Pg. 290-6 (Jan 2015) ISSN: 1530-8561 [Electronic] England
• PMID: 25361948 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: © 2014 American Association for Clinical Chemistry.

Chemical References

• DNA, Circular
• DNA, Viral
• Hepatitis B Surface Antigens

Topics

• Carcinoma, Hepatocellular (pathology, virology)
• DNA, Circular (genetics)
• DNA, Viral (genetics)
• Female
• Gene Dosage
• Hepatitis B (pathology, virology)
• Hepatitis B Surface Antigens (blood, genetics)
• Hepatitis B virus (genetics, isolation & purification)
• Humans
• Liver Neoplasms (pathology, virology)
• Male
• Middle Aged
• Neoplasm Staging
• Paraffin Embedding
• Polymerase Chain Reaction (methods)