Direct Detection of Drug-Resistant Hepatitis B Virus in Serum Using a Dendron-Modified Microarray
0
조회
0
다운로드
0
인용
0
참고문헌
키워드
초록
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, laborintensive, 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/L). 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 patie
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, laborintensive, 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/L). 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 patie
참고문헌 (35)
- Perz JF, "The contributions of hepatitis B virus and hepatitis C virus infections to cirrhosis and primary liver cancer worldwide", J Hepatol, 45, pp.529~538 (2006)[DOI]
- Schiff ER, "Prevention of mortality from hepatitis B and hepatitis C", Lancet, 368, pp.896~897 (2006)[DOI]
- Chen CJ, "Risk of hepatocellular carcinoma across a biological gradient of serum hepatitis B virus DNA level", JAMA, 295, pp.65~73 (2006)[DOI]
- Liaw YF, "Lamivudine for patients with chronic hepatitis B and advanced liver disease", N Engl J Med, 351, pp.1521~1531 (2004)[DOI]
- Hoofnagle JH, "Management of hepatitis B: summary of a clinical research workshop", Hepatology, 45, pp.1056~1075 (2007)[DOI]
- Zoulim F, "Hepatitis B virus resistance to nucleos(t)ide analogues", Gastroenterology, 137, pp.1593~1608 (2009)[DOI]
- Yim HJ, "Options for the management of antiviral resistance during hepatitis B therapy: reflections on battles over a decade", Clin Mol Hepatol, 19, pp.195~209 (2013)[DOI]
- Lee JH, "Prior exposure to lamivudine increases entecavir resistance risk in chronic hepatitis B Patients without detectable lamivudine resistance", Antimicrob Agents Chemother, 58, pp.1730~1737 (2014)[DOI]
- Ou ZY, "Rapid and accurate genotyping of YMDD motif variants in the hepatitis B virus genome by an improved reverse dot blot method", J Clin Microbiol, 43, pp.5685~5689 (2005)[DOI]
- Whalley SA, "Monitoring the emergence of hepatitis B virus polymerase gene variants during lamivudine therapy using the LightCycler", J Clin Microbiol, 39, pp.1456~1459 (2001)[DOI]
- Libbrecht E, "Evolution of primary and compensatory lamivudine resistance mutations in chronic hepatitis B virus-infected patients during long-term lamivudine treatment, assessed by a line probe assay", J Clin Microbiol, 45, pp.3935~3941 (2007)[DOI]
- Lindstrom A, "Pyrosequencing for detection of lamivudine-resistant hepatitis B virus", J Clin Microbiol, 42, pp.4788~4795 (2004)[DOI]
- Kirishima T, "Detection of YMDD mutant using a novel sensitive method in chronic liver disease type B patients before and during lamivudine treatment", J Hepatol, 37, pp.259~265 (2002)[DOI]
- Jang H, "Oligonucleotide chip for detection of Lamivudine-resistant hepatitis B virus", J Clin Microbiol, 42, pp.4181~4188 (2004)[DOI]
- Hong SP, "Detection of hepatitis B virus YMDD variants using mass spectrometric analysis of oligonucleotide fragments", J Hepatol, 40, pp.837~844 (2004)[DOI]
- Lee CH, "Predominance of hepatitis B virus YMDD mutants is prognostic of viral DNA breakthrough", Gastroenterology, 130, pp.1144~1152 (2006)[DOI]
- Kim JH, "Molecular diagnosis and treatment of drug-resistant hepatitis B virus", World J Gastroenterol, 20, pp.5708~5720 (2014)[DOI]
- Heo Jeong, "Detection of YMDD Motif Mutants by Oligonucleotide Chips in Lamivudine-Untreated Patients with Chronic Hepatitis B Virus Infection", Journal of Korean Medical Science, 19(4), pp.541~546 (2004), 대한의학회
- Vernet G, "The DNA-Chip technology as a new molecular tool for the detection of HBV mutants", J Clin Virol, 34(Suppl 1), pp.S49~S53 (2005)[DOI]
- Pas SD, "Comparison of reverse hybridization, microarray, and sequence analysis for genotyping hepatitis B virus", J Clin Microbiol, 46, pp.1268~1273 (2008)[DOI]
- Kwon SH, "DNA-DNA interaction on dendron-functionalized sol-gel silica films followed with surface plasmon fluorescence spectroscopy", J Colloid Interface Sci, 308, pp.325~331 (2007)[DOI]
- Hong BJ, "Nanoscale-controlled spacing provides DNA microarrays with the SNP discrimination efficiency in solution phase", Langmuir, 21, pp.4257~4261 (2005)[DOI]
- Hong BJ, "DNA microarrays on nanoscalecontrolled surface", Nucleic Acids Res, 33, pp.e106~ (2005)[DOI]
- Oh SJ, "DNA microarrays on a dendron-modified surface improve significantly the detection of single nucleotide variations in the p53 gene", Nucleic Acids Res, 33, pp.e90~ (2005)[DOI]
- Lim KH, "Suppression of interferon-mediated anti-HBV response by single CpG methylation in the 5'-UTR of TRIM22", Gut, 67, pp.166~178 (2018)[DOI]
- Park YK, "Cleaved c-FLIP mediates the antiviral effect of TNF-alpha against hepatitis B virus by dysregulating hepatocyte nuclear factors", J Hepatol, 64, pp.268~277 (2016)[DOI]
- Kim KT, "Dramatic increase in signal by integration of polymerase chain reaction and hybridization on surface of DNA microarray", Anal Biochem, 396, pp.139~145 (2010)[DOI]
- Tjong V, "Amplified on-chip fluorescence detection of DNA hybridization by surfaceinitiated enzymatic polymerization", Anal Chem, 83, pp.5153~5159 (2011)[DOI]
- Klein A, "Comparison of methods for extraction of nucleic acid from hemolytic serum for PCR amplification of hepatitis B virus DNA sequences", J Clin Microbiol, 35, pp.1897~1899 (1997)
- Kim JH, "Low-level viremia and the increased risk of hepatocellular carcinoma in patients receiving entecavir treatment", Hepatology, 66, pp.335~343 (2017)[DOI]
- Caligiuri P, "Overview of hepatitis B virus mutations and their implications in the management of infection", World J Gastroenterol, 22, pp.145~154 (2016)[DOI]
- Kim KH, "Discovery and development of anti-HBV agents and their resistance", Molecules, 15, pp.5878~5908 (2010)[DOI]
- Ahn SH, "The impact of the hepatitis B virus polymerase rtA181T mutation on replication and drug resistance is potentially affected by overlapping changes in surface gene", J Virol, 88, pp.6805~6818 (2014)[DOI]
- Corsa AC, "No resistance to tenofovir disoproxil fumarate through 96 weeks of treatment in patients with lamivudine-resistant chronic hepatitis B", Clin Gastroenterol Hepatol, 12, pp.2106~2112 (2014)[DOI]
- 정고은, "Tenofovir has inferior efficacy in adefovir-experienced chronic hepatitis B patients compared to nucleos(t)idenaïve patients", Clinical and Molecular Hepatology, 23(1), pp.66~73 (2017), 대한간학회[DOI]
