Gut Liver.
2018 May;12(3):331-341. 10.5009/gnl17336.
Direct Detection of Drug-Resistant Hepatitis B Virus in Serum Using a Dendron-Modified Microarray
- Affiliations
-
- 1Department of Pharmacology, Center for Cancer Research and Diagnostic Medicine, IBST, Seoul, Korea. khkim10@kku.ac.kr
- 2Department of Environmental and Tropical Medicine, Konkuk University School of Medicine, Seoul, Korea.
- 3Department of Internal Medicine, Konkuk University School of Medicine, Seoul, Korea.
- 4Department of Internal Medicine and Liver Research Institute, Seoul National University Hospital, Seoul National University College of Medicine, Seoul, Korea.
- 5KU Open Innovation Center, Konkuk University, Seoul, Korea.
- 6Research Institute of Medical Sciences, Konkuk University, Seoul, Korea.
- KMID: 2411679
- DOI: http://doi.org/10.5009/gnl17336
Abstract
- BACKGROUND/AIMS
Direct sequencing is the gold standard for the detection of drug-resistance mutations in hepatitis B virus (HBV); however, this procedure is time-consuming, labor-intensive, and difficult to adapt to high-throughput screening. In this study, we aimed to develop a dendron-modified DNA microarray for the detection of genotypic resistance mutations and evaluate its efficiency.
METHODS
The specificity, sensitivity, and selectivity of dendron-modified slides for the detection of representative drug-resistance mutations were evaluated and compared to those of conventional slides. The diagnostic accuracy was validated using sera obtained from 13 patients who developed viral breakthrough during lamivudine, adefovir, or entecavir therapy and compared with the accuracy of restriction fragment mass polymorphism and direct sequencing data.
RESULTS
The dendron-modified slides significantly outperformed the conventional microarray slides and were able to detect HBV DNA at a very low level (1 copy/μL). Notably, HBV mutants could be detected in the chronic hepatitis B patient sera without virus purification. The validation of our data revealed that this technique is fully compatible with sequencing data of drug-resistant HBV.
CONCLUSIONS
We developed a novel diagnostic technique for the simultaneous detection of several drug-resistance mutations using a dendron-modified DNA microarray. This technique can be directly applied to sera from chronic hepatitis B patients who show resistance to several nucleos(t)ide analogues.
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