Retrospective Cohort Study
Copyright ©The Author(s) 2019. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Aug 7, 2019; 25(29): 3985-3995
Published online Aug 7, 2019. doi: 10.3748/wjg.v25.i29.3985
Liquid biopsy for non-invasive assessment of liver injury in hepatitis B patients
Wen-Ying Xia, Li Gao, Er-Hei Dai, Dan Chen, Er-Fu Xie, Li Yang, Shi-Chang Zhang, Bing-Feng Zhang, Jian Xu, Shi-Yang Pan
Wen-Ying Xia, Li Gao, Dan Chen, Er-Fu Xie, Shi-Chang Zhang, Bing-Feng Zhang, Jian Xu, Shi-Yang Pan, Department of Laboratory Medicine, the First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, Jiangsu Province, China
Er-Hei Dai, Li Yang, Department of Laboratory Medicine, the Fifth Hospital of Shijiazhuang, Shijiazhuang 050021, Hebei Province, China
Author contributions: Xia WY, Gao L, and Dai EH contributed equally to this work as co-first authors and finished the major experiments; Pan SY designed the research; Chen D, Xie EF, and Yang L collected and analyzed the data; Gao L and Zhang SC wrote the paper; Zhang BF and Xu J critically revised the manuscript for important content.
Supported by the National Natural Science Foundation of China, No. 81672100 and No. 81371894; the Key Laboratory for Laboratory Medicine of Jiangsu Province of China, No. ZDXKB2016005; and Jiangsu Provincial Commission of Health and Family Planning, No. H201609.
Institutional review board statement: This study was approved for publication by our Institutional Reviewer.
Informed consent statement: All study participants or their legal guardian provided informed written consent for personal and medical data collection prior to study enrolment.
Conflict-of-interest statement: All authors have no conflict of interest related to the manuscript.
Data sharing statement: The original anonymous dataset is available on request from the corresponding author at sypan@njmu.edu.cn.
STROBE statement: The authors have read the STROBE Statement-checklist of items, and the manuscript was prepared and revised according to STROBE Statement-checklist of items.
Open-Access: This article is an open-access article which was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: http://creativecommons.org/licenses/by-nc/4.0/
Corresponding author: Shi-Yang Pan, MD, Professor, Department of Laboratory Medicine, the First Affiliated Hospital of Nanjing Medical University, No. 300, Guangzhou Road, Nanjing 210029, Jiangsu Province, China. sypan@njmu.edu.cn
Telephone: +86-25-68303452
Received: March 26, 2019
Peer-review started: March 26, 2019
First decision: May 24, 2019
Revised: June 13, 2019
Accepted: July 5, 2019
Article in press: July 3, 2019
Published online: August 7, 2019
Processing time: 134 Days and 6.4 Hours
Abstract
BACKGROUND

Hepatitis B is a major public health problem in China. Accurate liver injury assessment is essential for clinical evidence-based treatment. Liver biopsy is considered the gold standard method to stage liver disease, but it is not widely used in resource-limited settings. Therefore, non-invasive liquid biopsy tests are needed.

AIM

To assess liver injury in hepatitis B patients using quantified cell free DNA combined with other serum biomarker as a liquid biopsy-based method.

METHODS

A cohort of 663 subjects including 313 hepatitis B patients and 350 healthy controls were enrolled. Ultrasound-guided liver biopsies followed by histopathological assessments were performed for the 263 chronic hepatitis B patients to determine the degree of liver injury. Cell-free DNA was quantified using a novel duplex real-time polymerase chain reaction assay.

RESULTS

Compared with healthy controls, patients with hepatitis B virus (HBV) infection had significantly higher plasma DNA, serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), bilirubin, and HBV DNA levels (P < 0.01). Serum ALT, AST, bilirubin, and plasma DNA levels of patients with marked-severe inflammation were significantly higher than those with mild-moderate inflammation (P < 0.01). There was a statistically significant correlation between hepatocyte inflammation severity and serum bilirubin (R2 = 0.673, P < 0.01) or plasma DNA (R2 = 0.597, P < 0.01) levels. The areas under the curves of serum ALT, bilirubin, plasma DNA, and their combination to distinguish between patients with mild–moderate and marked-severe inflammation were 0.8059, 0.7910, 0.7921, and 0.9564, respectively.

CONCLUSION

The combination of plasma DNA, serum ALT, and bilirubin could be a candidate liquid biopsy for non-invasive assessment of liver injury in hepatitis B patients.

Keywords: Liquid biopsy; plasma DNA; Hepatitis B; Alanine aminotransferase; Duplex real-time quantitative polymerase chain reaction

Core tip: Our study used quantified cell free DNA combined with other serum biomarker as a liquid biopsy-based method to assess liver injury in hepatitis B patients. A cohort of 663 subjects including 313 hepatitis B patients and 350 healthy controls were enrolled. Ultrasound-guided liver biopsies followed by histopathological assessments were performed for the 263 chronic hepatitis B patients to determine the degree of liver injury. Cell-free DNA was quantified using a novel duplex real-time polymerase chain reaction assay. Our results demonstrated that the combination of plasma DNA, serum alanine aminotransferase, and bilirubin could be a candidate liquid biopsy for non-invasive assessment of liver injury in hepatitis B patients.