Application of in situ hybridization in the detection of hepatitis B virus nucleic acids and cccDNA
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摘要: 原位杂交技术(ISH)是分子生物学、组织化学及细胞学相结合而产生的一门新兴技术,能够在细胞和染色体水平上对特定核酸进行定量与定位,广泛应用于病毒学研究。ISH对于HBV中的核酸(RNA、复制中间体DNA),以及共价闭合环状DNA的检测有着重要意义,就这一技术的发展以及在HBV研究中的应用作梳理与总结。Abstract: In situ hybridization (ISH) is a new technique which combines molecular biology, histochemistry, and cytology. It can quantify and locate specific nucleic acids at the cellular and chromosomal levels and is widely used in virological research. ISH is of great significance for the detection of hepatitis B virus (HBV) nucleic acids (RNA and replicative intermediate DNA) and covalently closed circular DNA.This article reviews the development of ISH and its application in HBV research.
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Key words:
- hepatitis B virus /
- cccDNA /
- in situ hybridization
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[1]REVILL PA, CHISARI FV, BLOCK JM, et al.A global scientific strategy to cure hepatitis B[J].Lancet Gastroenterol Hepatol, 2019.[Epub ahead of print] [2]YAN H, ZHONG G, XU G, et al.Sodium taurocholate cotransporting polypeptide is a functional receptor for human hepatitis B and D virus[J].Elife, 2012, 1:e00049. [3]GUO JT, GUO H.Metabolism and function of hepatitis B virus cccDNA:Implications for the development of cccDNA-targeting antiviral therapeutics[J].Antiviral Res, 2015, 122:91-100. [4]SEEGER C, MASON WS.Hepatitis B virus biology[J].Microbiol Mol Biol Rev, 2000, 64 (1) :51-68. [5]PERRILLO R.Benefits and risks of interferon therapy for hepatitis B[J].Hepatology, 2009, 49 (5 Suppl) :s103-s111. [6]DURANTEL D, ZOULIM F.New antiviral targets for innovative treatment concepts for hepatitis B virus and hepatitis delta virus[J].J Hepatol, 2016, 64 (1 Suppl) :s117-s131. [7]CHEN Y, SZE J, HE ML.HBV cccDNA in patients'sera as an indicator for HBV reactivation and an early signal of liver damage[J].World J Gastroenterol, 2004, 10 (1) :82-85. [8]YU XH, WANG Y, YANG TT.Research advances in clinical application of hepatitis B virus covalently closed circular DNA[J].Chin J Nosocomiol, 2013, 23 (17) :4338-4340. (in Chinese) 余晓红, 王媛, 杨廷桐.乙型肝炎病毒cccDNA临床应用研究新进展[J].中华医院感染学杂志, 2013, 23 (17) :4338-4340. [9]TUTTLEMAN JS, POURCEL C, SUMMERS J.Formation of the pool of covalently closed circular viral DNA in hepadnavirusinfected cells[J].Cell, 1986, 47 (3) :451-460. [10]ZHANG YY, ZHANG B, THEELE D, et al.Single-cell analysis of covalently closed circular DNA copy numbers in a hepadnavirus-infected liver[J].Proc Natl Acad Sci U S A, 2003, 100 (21) :12372-12377. [11]DANDRI M, BURDA MR, WILL H, et al.Increased hepatocyte turnover and inhibition of woodchuck hepatitis B virus replication by adefovir in vitro do not lead to reduction of the closed circular DNA[J].Hepatology, 2000, 32 (1) :139-146. [12] WERLE-LAPOSTOLLE B, BOWDEN S, LOCARNINI S, et al.Persistence of cccDNA during the natural history of chronic hepatitis B and decline during adefovir dipivoxil therapy[J].Gastroenterology, 2004, 126 (7) :1750-1758. [13]NASSAL M.HBV cccDNA:Viral persistence reservoir and key obstacle for a cure of chronic hepatitis B[J].Gut, 2015, 64 (12) :1972-1984. [14]ITZKOVITZ S, van OUDENAARDEN A.Validating transcripts with probes and imaging technology[J].Nat Methods, 2011, 8 (4Suppl) :s12-s19. [15]GALL JG, PARDUE ML.Formation and detection of RNA-DNAhybrid molecules in cytological preparations[J].Proc Natl Acad Sci U S A, 1969, 63 (2) :378-383. [16]JOHN HA, BIRNSTIEL ML, JONES KW.RNA-DNA hybrids at the cytological level[J].Nature, 1969, 223 (5206) :582-587. [17]BAUMAN JG, WIEGANT J, BORST P, et al.A new method for fluorescence microscopical localization of specific DNA sequences by in situ hybridization of fluorochromelabelled RNA[J].Exp Cell Res, 1980, 128 (2) :485-490. [18]LANGER PR, WALDROP AA, WARD DC.Enzymatic synthesis of biotin-labeled polynucleotides:Novel nucleic acid affinity probes[J].Proc Natl Acad Sci U S A, 1981, 78 (11) :6633-6637. [19]NAOUMOV NV, ALEXANDER GJ, EDDLESTON AL, et al.In situ hybridisation in formalin fixed, paraffin wax embedded liver specimens:Method for detecting human and viral DNA using biotinylated probes[J].J Clin Pathol, 1988, 41 (7) :793-798. [20]HEILES HB, GENERSCH E, KESSLER C, et al.In situ hybridization with digoxigenin-labeled DNA of human papillomaviruses (HPV 16/18) in He La and Si Ha cells[J].Biotechniques, 1988, 6 (10) :978-981. [21]HAASE AT, RETZEL EF, STASKUS KA.Amplification and detection of lentiviral DNA inside cells[J].Proc Natl Acad Sci US A, 1990, 87 (13) :4971-4975. [22]KERN D, COLLINS M, FULTZ T, et al.An enhanced-sensitivity branched-DNA assay for quantification of human immunodeficiency virus type 1 RNA in plasma[J].J Clin Microbiol, 1996, 34 (12) :3196-3202. [23]HENDRICKS DA, STOWE BJ, HOO BS, et al.Quantitation of HBV DNA in human serum using a branched DNA (bDNA) signal amplification assay[J].Am J Clin Pathol, 1995, 104 (5) :537-546. [24]DETMER J, LAGIER R, FLYNN J, et al.Accurate quantification of hepatitis C virus (HCV) RNA from all HCV genotypes by using branched-DNA technology[J].J Clin Microbiol, 1996, 34 (4) :901-907. [25]PLAYER AN, SHEN LP, KENNY D, et al.Single-copy gene detection using branched DNA (bDNA) in situ hybridization[J].J Histochem Cytochem, 2001, 49 (5) :603-612. [26]REN JY, CHEN ML, TIAN GS, et al.Application of in situ hybridization in research on hepatitis B[J].Foreign Med Sci:Epidemiol Loimol, 1988, 6:245-248. (in Chinese) 任进余, 陈梅龄, 田庚善, 等.原位杂交技术在乙型肝炎研究中的应用[J].国外医学.流行病学传染病学分册, 1988, 6:245-248. [27]GOWANS EJ, BURRELL CJ, JILBERT AR, et al.Detection of hepatitis B virus DNA sequences in infected hepatocytes by in situ cytohybridisation[J].J Med Virol, 1981, 8 (1) :67-78. [28]NURIYA H, INOUE K, TANAKA T, et al.Detection of hepatitis B and C viruses in almost all hepatocytes by modified PCR-based in situ hybridization[J].J Clin Microbiol, 2010, 48 (11) :3843-3851. [29]KENNY D, SHEN LP, KOLBERG JA.Detection of viral infection and gene expression in clinical tissue specimens using branched DNA (bDNA) in situ hybridization[J].J Histochem Cytochem, 2002, 50 (9) :1219-1227. [30]CALABRESE D, WIELAND SF.Highly sensitive detection of HBVRNA in liver tissue by in situ hybridization[J].Methods Mol Biol, 2017, 1540:119-134. [31]YEH CT, CHIU HT, CHU CM, et al.G1 phase dependent nuclear localization of relaxed-circular hepatitis B virus DNA and aphidicolin-induced accumulation of covalently closed circular DNA[J].J Med Virol, 1998, 55 (1) :42-50. [32]ZHONG Y, HU S, XU C, et al.A novel method for detection of HBVcccDNA in hepatocytes using rolling circle amplification combined with in situ PCR[J].BMC Infect Dis, 2014, 14:608. [33]ZHANG X, LU W, ZHENG Y, et al.In situ analysis of intrahepatic virological events in chronic hepatitis B virus infection[J].J Clin Invest, 2016, 126 (3) :1079-1092. [34]LI M, SOHN JA, SEEGER C.Distribution of hepatitis B virus nuclear DNA[J].J Virol, 2018, 92 (1) :e01391-17. [35]ZHANG X, YUE L, ZHANG Z, et al.Establishment of a fluorescent in situ hybridization assay for imaging hepatitis B virus nucleic acids in cell culture models[J].Emerg Microbes Infect, 2017, 6 (11) :e98.
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