HBV cccDNA定量检测方法的研究进展

江培学; 毛日成; 张继明

doi:10.3969/j.issn.1001-5256.2019.06.006

HBV cccDNA定量检测方法的研究进展

DOI: 10.3969/j.issn.1001-5256.2019.06.006

详细信息

中图分类号: R512.62
计量
- 文章访问数: 2764
- HTML全文浏览量: 42
- PDF下载量: 465
- 被引次数: 0
出版历程
- 收稿日期: 2019-05-09
- 出版日期: 2019-06-20

Recent advances of quantitative detection methods for HBV cccDNA

摘要

摘要: 从慢性HBV感染的肝细胞中清除HBV cccDNA被认为是根除HBV的关键。在抗病毒治疗之前、期间和之后监测HBV cccDNA对于慢性乙型肝炎患者的疗效评估极为重要;随着靶向HBV cccDNA的抗HBV新药研发的不断推进,也迫切需要准确而灵敏的HBV cccDNA检测方法,以评价其疗效。近年来,为了提高HBV cccDNA定量检测的特异度,在传统的PCR定量检测方法的基础上进行了改进。同时,为了提高敏感度,还推出了HBV cccDNA的数字PCR定量检测方法等。本文综述了引入酶切的qPCR法、磁珠捕获杂交法、滚环扩增结合原位PCR法、数字PCR法和单细胞内数字PCR法等方法在HBV cccDNA定量检测方面的应用进展。
- 乙型肝炎病毒 /
- 共价闭合环状DNA /
- 检测方法
Abstract: Elimination of HBV cccDNA from hepatocytes infected with chronic HBV virus is considered to be the key to eradicating HBV.Monitoring HBV cccDNA before, during, and after viral treatment is essential for routine treatment of patients with chronic hepatitis B. With the introduction of new anti-HBV treatment technologies and new drugs targeting HBV cccDNA, Accurate and sensitive HBV cccDNA assays are urgently needed to evaluate efficacy. In recent years, HBV cccDNA detection methods have achieved gratifying results in both traditional PCR methods and digital PCR methods popular in recent years. In this paper, the advances in HBV cccDNA quantitative detection by qPCR, Magnetic bead capture hybridization, rolling circle amplification combined with in situ PCR, digital PCR and digital PCR assay in single cells were reviewed.
- hepatitis B virus /
- cccDNA /
- quantitative detection method

HTML全文

参考文献(36)

[1]RAZAVI-SHEARER D, GAMKRELIDZE I, NGUYEN MH, et al.Global prevalence, treatment, and prevention of hepatitis B virus infection in 2016:A modelling study[J].Lancet Gastroenterol Hepatol, 2018, 3 (6) :383-403.

[2]CHEN J, WU M, LIU K, et al.New insights into hepatitis B virus biology and implications for novel antiviral strategies[J].Nat Sci Rev, 2015, 2 (4) :296-313.

[3]SEEGER C, MASON WS.Molecular biology of hepatitis B virus infection[J].Virology, 2015, 8 (12) :672-686.

[4]LI XL, ZHAO JH, YUAN Q, et al.Detection of HBV covalently closed circular DNA[J].Viruses, 2017, 9 (3) :139-158.

[5]CAI DW, NIE H, YAN R, et al.A Southern blot assay for detection of hepatitis B virus covalently closed circular DNA from cell cultures[J].Methods Mol Biol, 2013, 1030:151-161.

[6]KOCK J, SCHLICHT HG.Analysis of the earliest steps of hepadnavirus replication:Genome repair after infectious entry into hepatocytes does not depend on viral polymerase activity[J].J Virol, 1993, 67 (11) :4867-4874.

[7]ADDISON WR, WONG WW, FISCHER KP, et al.A quantitative competitive PCR assay for the covalently closed circular form of the duck hepatitis B virus[J].Antiviral Res, 2000, 48 (7) :27-37.

[8]HE ML, WU J, CHEN Y, et al.A new and sensitive method for the quantification of HBV cccDNA by real-time PCR[J].Biochem Biophys Res Commun, 2002, 295:1102-1107.

[9]SHAO JB, CHEN Z, NI WQ, et al.A quantitative method to detect HBV cccDNA by chimeric primer and real-time polymerase chain reaction[J].J Virol Method, 2003, 112:45-52.

[10]WONG DK, YUEN MF, YUAN H, et al.Quantitation of covalently closed circular hepatitis B virus DNA in chronic hepatitis B patients[J].Hepatology, 2004, 40 (3) :727-737.

[11]WERLE-LAPOSTOLLE B, BOWDEN S, ZOULIM F, et al.Persistence of HBV cccDNA during the natural history of chronic hepatitis B and decline during adefovir dipivoxil therapy[J].Gastroenterology, 2004, 126 (11) :1750-1758.

[12]NASSAL M.HBV cccDNA:Viral persistence reservoir and key obstacle for a cure of chronic hepatitis B[J].Gut, 2015, 34 (1) :1-13.

[13]LUO J, CUI X, HU J.Identification of intermediate in hepatitis B virus cccDNA.formation and sensitive and selective cccD-NA detection[J].J Virol, 2017, 8 (3) :539-556.

[14]JIANG PX, MAO RC, ZHANG JM, et al.Exonuclease I and IIIimprove the detection efficacy of hepatitis B virus covalently closed circular DNA[J].Hepatobiliary Pancreat Dis Int, 2018, 19 (18) :261-269.

[15]QU B, NI Y, LEMPP FA, et al.T5 exonuclease hydrolysis of hepatitis B virus replicative intermediates allows reliable quantification and fast drug efficacy testing of covalently closed circular DNA by PCR[J].J Virol, 2018, 92 (23) :e01117-e01118.

[16]YEH CT, CHIU HT, CHU CM.G1 phase dependent nuclear localization of rcDNA and aphindicolin induced HBV cccDNA[J].J Med Virol, 1998, 55 (5) :42-50.

[17]GUO YC, SHENG SC, NIE B, et al.Development of magnetic capture hybridization and quantitative polymerase chain reaction for hepatitis B virus covalently closed circular DNA[J].Hepat Mon, 2015, 15 (6) :729-736.

[18]MARGERIDON S, CARROUE DS, CHEMIN I, et al.Rolling circle amplification, a powerful tool for genetic and functional studies of complete hepatitis B virus genomes from low-level infections and for directly probing covalently closed circular DNA[J].Antimicrob Agents Chemother, 2008, 52 (9) :3068-3073.

[19]ZHONG YW, HAN JQ, ZOU ZS, et al.Quantitation of HBV covalently closed circular DNA in micro formalin fixed paraffinembedded liver tissue using rolling circle amplification in combination with real-time PCR[J].Clin Chim Acta, 2011, 412 (6) :1905-1911.

[20]ZHONG YW, HU SY, XU C, et al.A novel method for detection of HBV cccDNA in hepatocytes using rolling circle amplification combined with in situ PCR[J].BMC infect Dis, 2014, 14:608-619.

[21]ZHANG XN, LU W, ZHENG Y, et al.In situ analysis of intrahepatic virological events in chronic hepatitis B virus infection[J].J Clin Investig, 2016, 126 (2) :1079-1092.

[22]WHITE RA, QUAKE SR, CURR K.Digital PCR provides absolute quantitation of viral load for an occult RNA virus[J].J Virol Methods, 2011, 179 (6) :45-50.

[23]HINDSON BJ, NESS KD, MASQUELIER DA, et al.Highthroughput droplet digital PCR system for absolute quantitation of DNA copy number[J].Anal Chem, 2011, 83 (12) :8604-8610.

[24]VOGELSTEIN B, KINZLER KW.Digital PCR[J].Proc Natl Acad Sci U S A, 1999, 96 (4) :9236-9241.

[25]HAYDEN RT, GU Z, INGERSOLL J.Comparison of droplet digital PCR to real-time PCR for quantitative detection of cytomegalovirus[J].J Clin Microbiol, 2013, 51 (11) :540-546.

[26]MU D, YAN L, TANG H, et al.A sensitive and accurate quantification method for the detection of hepatitis B virus covalently closed circular DNA by the application of a droplet digital polymerase chain reaction amplification system[J].Biotechnol Lett, 2015, 37 (10) :2063-2073.

[27]CAVIGLIA GP, ABATE ML, TANDOI F, et al.Quantitation of HBV cccDNA in anti-HBc-positive liver donors by droplet digital PCR:A new tool to detect occult infection[J].J Hepatol, 2018, 69 (2) :301-307.

[28]WALLRAPP A, RIESENFELD SJ, BURKETT PR, et al.The neuropeptide NMU amplifies ILC2-driven allergic lung inflammation[J].Nature, 2017, 549 (7672) :351-356.

[29]YILMAZ S, SINGH AK.Single cell genome sequencing[J].Curr Opin Biotechnol, 2012, 23 (3) :437-443.

[30]ZONG CZ, LU SJ, CHAPMAN AR, et al.Genome-wide detection of single nucleotide and copy number variations of a single human Cell[J].Science, 2012, 338 (6114) :1622-1626.

[31]LU SJ, ZONG CZ, XIE XS, et al.Probing meiotic recombination and aneuploidy of single sperm cells by whole genome sequencing using MALBAC[J].Science, 2012, 338 (6114) :1627-1630.

[32]HUANG JT, YANG Y, LIU SM, et al.A highly sensitive and robust method for hepatitis B virus covalently closed circular DNA detection in single cells and serum[J].J Mol, 2018, 20 (3) :334-343.

[33]ZOULIM F.New insight on hepatitis B virus persistence from the study of intrahepatic viral HBV cccDNA[J].J Hepatol, 2005, 42 (3) :302-308.

[34]LUCIFORA J, SALVETTI A, MARNIQUET X, et al.Detection of the hepatitis B virus (HBV) covalently-closed-circular DNA (HBV cccDNA) in mice transduced with a recombinant AAV-HBV vector[J].Antiviral Res, 2017, 145 (9) :14-19.

[35]YUEN MF, WONG DK, SUM SS, et al.Effect of lamivudine therapy on the serum covalently closed-circular (ccc) DNAof chronic hepatitis B infection[J].Am J Gastroenterol, 2005, 100 (5) :1099-1103.

[36]CHEN YJ, 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.

施引文献

资源附件(0)

访问统计