{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,12]],"date-time":"2026-10-12T20:21:29Z","timestamp":1791836489944,"version":"4.3.4"},"reference-count":33,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T00:00:00Z","timestamp":1760054400000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0"},{"start":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T00:00:00Z","timestamp":1760054400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0"}],"funder":[{"DOI":"10.13039\/501100012165","name":"Key Technologies Research and Development Program","doi-asserted-by":"publisher","award":["2023YFC2307600"],"award-info":[{"award-number":["2023YFC2307600"]}],"id":[{"id":"10.13039\/501100012165","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100005147","name":"Applied Basic Research Key Project of Yunnan","doi-asserted-by":"publisher","award":["202401BF070001-019"],"award-info":[{"award-number":["202401BF070001-019"]}],"id":[{"id":"10.13039\/501100005147","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100010905","name":"Major Research Plan","doi-asserted-by":"publisher","award":["32371000"],"award-info":[{"award-number":["32371000"]}],"id":[{"id":"10.13039\/501100010905","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100019511","name":"Joint Fund to Promote Cross-Straits Scientific and Technological Cooperation","doi-asserted-by":"publisher","award":["82060368"],"award-info":[{"award-number":["82060368"]}],"id":[{"id":"10.13039\/501100019511","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["J Transl Med"],"DOI":"10.1186\/s12967-025-07131-9","type":"journal-article","created":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T08:13:14Z","timestamp":1760083994000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["A single-cell digital PCR method tailored for quantification of HBV DNA positive cells in liver biopsy tissues"],"prefix":"10.1186","volume":"23","author":[{"given":"Jingmei","family":"Li","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hui","family":"Li","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xin","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xueyan","family":"Liu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zumi","family":"Zhou","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jia","family":"Wei","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rujuan","family":"Zhao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tai-Cheng","family":"Zhou","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zijie","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8227-9719","authenticated-orcid":false,"given":"Chunmei","family":"Li","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2025,10,10]]},"reference":[{"key":"7131_CR1","unstructured":"World Health Organization. Hepatitis B. [Internet] 2024 9 April 2024 [cited 2025 28 May]; Available from: https:\/\/www.who.int\/news-room\/fact-sheets\/detail\/hepatitis-b."},{"issue":"12","key":"7131_CR2","doi-asserted-by":"publisher","first-page":"2354","DOI":"10.1136\/gutjnl-2023-330691","volume":"72","author":"ZQ Liu","year":"2023","unstructured":"Liu ZQ, Lin CQ, Mao XH, Guo CN, Suo C, Zhu DL, et al. Changing prevalence of chronic hepatitis B virus infection in China between 1973 and 2021: a systematic literature review and meta-analysis of 3740 studies and 231 million people. Gut. 2023;72(12):2354\u201363.","journal-title":"Gut"},{"issue":"3","key":"7131_CR3","doi-asserted-by":"publisher","first-page":"230","DOI":"10.2471\/BLT.18.219469","volume":"97","author":"J Liu","year":"2019","unstructured":"Liu J, Liang WN, Jing WZ, Liu M. Countdown to 2030: eliminating hepatitis B disease, China. Bull World Health Organ. 2019;97(3):230\u20138.","journal-title":"Bull World Health Organ"},{"issue":"1","key":"7131_CR4","doi-asserted-by":"publisher","first-page":"852","DOI":"10.1080\/22221751.2021.1919034","volume":"10","author":"J Hu","year":"2021","unstructured":"Hu J, Tang LD, Cheng JJ, Zhou TL, Li YH, Chang JH, et al. Hepatitis B virus nucleocapsid uncoating: biological consequences and regulation by cellular nucleases. Emerg Microb Infect. 2021;10(1):852\u201364.","journal-title":"Emerg Microb Infect"},{"issue":"3","key":"7131_CR5","doi-asserted-by":"publisher","first-page":"706","DOI":"10.1016\/j.jhep.2021.05.013","volume":"75","author":"MG Martinez","year":"2021","unstructured":"Martinez MG, Boyd A, Combe E, Testoni B, Zoulim F. Covalently closed circular DNA: the ultimate therapeutic target for curing HBV infections. J Hepatol. 2021;75(3):706\u201317.","journal-title":"J Hepatol"},{"issue":"5","key":"7131_CR6","doi-asserted-by":"publisher","first-page":"972","DOI":"10.1136\/gutjnl-2022-328380","volume":"72","author":"L Allweiss","year":"2023","unstructured":"Allweiss L, Testoni B, Yu M, Lucifora J, Ko C, Qu BQ, et al. Quantification of the hepatitis B virus cccDNA: evidence-based guidelines for monitoring the key obstacle of HBV cure. Gut. 2023;72(5):972\u201383.","journal-title":"Gut"},{"issue":"11","key":"7131_CR7","doi-asserted-by":"publisher","first-page":"727","DOI":"10.1038\/s41575-022-00649-z","volume":"19","author":"A Kramvis","year":"2022","unstructured":"Kramvis A, Chang KM, Dandri M, Farci P, Glebe D, Hu JM, et al. A roadmap for serum biomarkers for hepatitis B virus: current status and future outlook. Nat Rev Gastroenterol Hepatol. 2022;19(11):727\u201345.","journal-title":"Nat Rev Gastroenterol Hepatol"},{"issue":"3","key":"7131_CR8","doi-asserted-by":"publisher","first-page":"539","DOI":"10.1016\/j.jhep.2019.11.003","volume":"72","author":"M Cornberg","year":"2020","unstructured":"Cornberg M, Lok ASF, Terrault NA, Zoulim F, Berg T, Brunetto MR, et al. Guidance for design and endpoints of clinical trials in chronic hepatitis B - Report from the 2019 EASL-AASLD HBV Treatment Endpoints Conferences. J Hepatol. 2020;72(3):539\u201357.","journal-title":"J Hepatol"},{"issue":"2","key":"7131_CR9","doi-asserted-by":"publisher","first-page":"275","DOI":"10.1016\/j.jhep.2016.08.022","volume":"66","author":"CL Lai","year":"2017","unstructured":"Lai CL, Wong D, Ip P, Kopaniszen M, Seto WK, Fung J, et al. Reduction of covalently closed circular DNA with long-term nucleos(t)ide analogue treatment in chronic hepatitis B. J Hepatol. 2017;66(2):275\u201381.","journal-title":"J Hepatol"},{"issue":"1","key":"7131_CR10","doi-asserted-by":"publisher","first-page":"2339944","DOI":"10.1080\/22221751.2024.2339944","volume":"13","author":"J Wang","year":"2024","unstructured":"Wang J, Zhu L, Zhang S, Zhang Z, Fan T, Cao F, et al. Clinical outcomes of treatment-na\u00efve HBeAg-negative patients with chronic hepatitis B virus infection with low serum HBsAg and undetectable HBV DNA. Emerg Microbes Infect. 2024;13(1):2339944.","journal-title":"Emerg Microbes Infect"},{"issue":"2","key":"7131_CR11","doi-asserted-by":"publisher","first-page":"409","DOI":"10.1016\/j.jhep.2020.04.013","volume":"73","author":"I Tout","year":"2020","unstructured":"Tout I, Loureiro D, Mansouri A, Soumelis V, Boyer N, Asselah T. Hepatitis B surface antigen seroclearance: immune mechanisms, clinical impact, importance for drug development. J Hepatol. 2020;73(2):409\u201322.","journal-title":"J Hepatol"},{"issue":"1","key":"7131_CR12","doi-asserted-by":"publisher","first-page":"42","DOI":"10.1002\/hep.31026","volume":"72","author":"I Carey","year":"2020","unstructured":"Carey I, Gersch J, Wang B, Moigboi C, Kuhns M, Cloherty G, et al. Pregenomic HBV RNA and Hepatitis B core-related antigen predict outcomes in Hepatitis B e antigen-negative chronic hepatitis B patients suppressed on nucleos(t)ide analogue therapy. Hepatology. 2020;72(1):42\u201357.","journal-title":"Hepatology"},{"issue":"1","key":"7131_CR13","doi-asserted-by":"publisher","first-page":"43","DOI":"10.1111\/apt.14376","volume":"47","author":"LY Mak","year":"2018","unstructured":"Mak LY, Wong DKH, Cheung KS, Seto WK, Lai CL, Yuen MF. Review article: hepatitis B core-related antigen (HBcrAg): an emerging marker for chronic hepatitis B virus infection. Aliment Pharmacol Ther. 2018;47(1):43\u201354.","journal-title":"Aliment Pharmacol Ther"},{"issue":"4","key":"7131_CR14","doi-asserted-by":"publisher","first-page":"1816","DOI":"10.1002\/hep.30325","volume":"69","author":"S Liu","year":"2019","unstructured":"Liu S, Zhou B, Valdes JD, Sun J, Guo HT. Serum hepatitis B virus RNA: a new potential biomarker for chronic hepatitis B virus infection. Hepatology. 2019;69(4):1816\u201327.","journal-title":"Hepatology"},{"key":"7131_CR15","doi-asserted-by":"publisher","first-page":"151","DOI":"10.1007\/978-1-62703-484-5_13","volume":"1030","author":"D Cai","year":"2013","unstructured":"Cai D, Nie H, Yan R, Guo JT, Block TM, Guo H. A southern blot assay for detection of hepatitis B virus covalently closed circular DNA from cell cultures. Methods Mol Biol. 2013;1030:151\u201361.","journal-title":"Methods Mol Biol"},{"issue":"2","key":"7131_CR16","doi-asserted-by":"publisher","first-page":"301","DOI":"10.1016\/j.jhep.2018.03.021","volume":"69","author":"GP Caviglia","year":"2018","unstructured":"Caviglia GP, Abate ML, Tandoi F, Ciancio A, Amoroso A, Salizzoni M, et al. Quantitation of HBV cccDNA in anti-HBc-positive liver donors by droplet digital PCR: a new tool to detect occult infection. J Hepatol. 2018;69(2):301\u20137.","journal-title":"J Hepatol"},{"issue":"3","key":"7131_CR17","doi-asserted-by":"publisher","first-page":"843","DOI":"10.1053\/j.gastro.2007.06.057","volume":"133","author":"T Volz","year":"2007","unstructured":"Volz T, Lutgehetmann M, Wachtler P, Jacob A, Quaas A, Murray JM, et al. Impaired intrahepatic hepatitis B virus productivity contributes to low viremia in most HBeAg-negative patients. Gastroenterology. 2007;133(3):843\u201352.","journal-title":"Gastroenterology"},{"key":"7131_CR18","doi-asserted-by":"publisher","first-page":"114344","DOI":"10.1016\/j.bios.2022.114344","volume":"211","author":"L Zhang","year":"2022","unstructured":"Zhang L, Parvin R, Fan Q, Ye F. Emerging digital PCR technology in precision medicine. Biosens Bioelectron. 2022;211:114344.","journal-title":"Biosens Bioelectron"},{"issue":"1","key":"7131_CR19","doi-asserted-by":"publisher","first-page":"3544","DOI":"10.1038\/s41467-019-11531-z","volume":"10","author":"J Lin","year":"2019","unstructured":"Lin J, Jordi C, Son M, Van Phan H, Drayman N, Abasiyanik MF, et al. Ultra-sensitive digital quantification of proteins and mRNA in single cells. Nat Commun. 2019;10(1):3544.","journal-title":"Nat Commun"},{"key":"7131_CR20","first-page":"8","volume":"6","author":"Y Igarashi","year":"2017","unstructured":"Igarashi Y, Uchiyama T, Minegishi T, Takahashi S, Watanabe N, Kawai T, et al. Single cell-based vector tracing in patients with ADA-SCID treated with stem cell gene therapy. Mol Ther. 2017;6:8\u201316.","journal-title":"Mol Ther"},{"key":"7131_CR21","doi-asserted-by":"publisher","DOI":"10.3390\/bios14020064","author":"W Fang","year":"2024","unstructured":"Fang W, Liu X, Maiga M, Cao W, Mu Y, Yan Q, et al. Digital PCR for single-cell analysis. Biosensors. 2024. https:\/\/doi.org\/10.3390\/bios14020064.","journal-title":"Biosensors"},{"issue":"6","key":"7131_CR22","doi-asserted-by":"publisher","first-page":"1087","DOI":"10.1016\/j.jhep.2024.06.037","volume":"81","author":"F Zoulim","year":"2024","unstructured":"Zoulim F, Chen PJ, Dandri M, Kennedy PT, Seeger C. Hepatitis B virus DNA integration: implications for diagnostics, therapy, and outcome. J Hepatol. 2024;81(6):1087\u201399.","journal-title":"J Hepatol"},{"key":"7131_CR23","doi-asserted-by":"publisher","first-page":"3","DOI":"10.1038\/s41378-023-00631-y","volume":"10","author":"T Tang","year":"2024","unstructured":"Tang T, Zhao H, Shen S, Yang L, Lim CT. Enhancing single-cell encapsulation in droplet microfluidics with fine-tunable on-chip sample enrichment. Microsyst Nanoeng. 2024;10:3.","journal-title":"Microsyst Nanoeng"},{"issue":"17","key":"7131_CR24","doi-asserted-by":"publisher","first-page":"3439","DOI":"10.1039\/C5LC00614G","volume":"15","author":"DJ Collins","year":"2015","unstructured":"Collins DJ, Neild A, deMello A, Liu AQ, Ai Y. The poisson distribution and beyond: methods for microfluidic droplet production and single cell encapsulation. Lab Chip. 2015;15(17):3439\u201359.","journal-title":"Lab Chip"},{"issue":"3","key":"7131_CR25","doi-asserted-by":"publisher","first-page":"433","DOI":"10.1080\/07388551.2022.2037503","volume":"43","author":"LL Tan","year":"2023","unstructured":"Tan LL, Loganathan N, Agarwalla S, Yang C, Yuan W, Zeng J, et al. Current commercial dPCR platforms: technology and market review. Crit Rev Biotechnol. 2023;43(3):433\u201364.","journal-title":"Crit Rev Biotechnol"},{"key":"7131_CR26","doi-asserted-by":"publisher","DOI":"10.3390\/v13081463","author":"L Wei","year":"2021","unstructured":"Wei L, Ploss A. Mechanism of hepatitis B virus cccDNA formation. Viruses. 2021. https:\/\/doi.org\/10.3390\/v13081463.","journal-title":"Viruses"},{"key":"7131_CR27","doi-asserted-by":"publisher","first-page":"109","DOI":"10.1007\/978-981-13-9151-4_5","volume":"1179","author":"F Li","year":"2020","unstructured":"Li F, Wang Z, Hu F, Su L. Cell Culture Models and Animal Models for HBV Study. Adv Exp Med Biol. 2020;1179:109\u201335.","journal-title":"Adv Exp Med Biol"},{"key":"7131_CR28","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1016\/j.coviro.2014.04.005","volume":"8","author":"L Men\u00e9ndez-Arias","year":"2014","unstructured":"Men\u00e9ndez-Arias L, \u00c1lvarez M, Pacheco B. Nucleoside\/nucleotide analog inhibitors of hepatitis B virus polymerase: mechanism of action and resistance. Curr Opin Virol. 2014;8:1\u20139.","journal-title":"Curr Opin Virol"},{"key":"7131_CR29","doi-asserted-by":"publisher","DOI":"10.1038\/emi.2014.64","author":"HC Yang","year":"2014","unstructured":"Yang HC, Kao JH. Persistence of hepatitis B virus covalently closed circular DNA in hepatocytes: molecular mechanisms and clinical significance. Emerg Microbes Infect. 2014. https:\/\/doi.org\/10.1038\/emi.2014.64.","journal-title":"Emerg Microbes Infect"},{"issue":"21","key":"7131_CR30","doi-asserted-by":"publisher","first-page":"12372","DOI":"10.1073\/pnas.2033898100","volume":"100","author":"YY Zhang","year":"2003","unstructured":"Zhang YY, Zhang BH, Theele D, Litwin S, Toll E, Summers J. Single-cell analysis of covalently closed circular DNA copy numbers in a hepadnavirus-infected liver. Proc Natl Acad Sci USA. 2003;100(21):12372\u20137.","journal-title":"Proc Natl Acad Sci USA"},{"issue":"3","key":"7131_CR31","doi-asserted-by":"publisher","first-page":"334","DOI":"10.1016\/j.jmoldx.2018.01.010","volume":"20","author":"JT Huang","year":"2018","unstructured":"Huang JT, Yang Y, Hu YM, Liu XH, Liao MY, Morgan R, et al. A highly sensitive and robust method for hepatitis B virus covalently closed circular DNA detection in single cells and serum. J Mol Diagn. 2018;20(3):334\u201343.","journal-title":"J Mol Diagn"},{"key":"7131_CR32","doi-asserted-by":"publisher","first-page":"104865","DOI":"10.1016\/j.antiviral.2020.104865","volume":"181","author":"T Tu","year":"2020","unstructured":"Tu T, Zehnder B, Qu B, Ni Y, Main N, Allweiss L, et al. A novel method to precisely quantify hepatitis B virus covalently closed circular (ccc)DNA formation and maintenance. Antivir Res. 2020;181:104865.","journal-title":"Antivir Res"},{"issue":"1","key":"7131_CR33","doi-asserted-by":"publisher","first-page":"100","DOI":"10.1016\/j.chom.2010.06.014","volume":"8","author":"SH Speck","year":"2010","unstructured":"Speck SH, Ganem D. Viral latency and its regulation: lessons from the gamma-herpesviruses. Cell Host Microbe. 2010;8(1):100\u201315.","journal-title":"Cell Host Microbe"}],"container-title":["Journal of Translational Medicine"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1186\/s12967-025-07131-9.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/article\/10.1186\/s12967-025-07131-9","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1186\/s12967-025-07131-9.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,4,7]],"date-time":"2026-04-07T12:27:08Z","timestamp":1775564828000},"score":1,"resource":{"primary":{"URL":"https:\/\/link.springer.com\/10.1186\/s12967-025-07131-9"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,10,10]]},"references-count":33,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2025,12]]}},"alternative-id":["7131"],"URL":"https:\/\/doi.org\/10.1186\/s12967-025-07131-9","relation":{},"ISSN":["1479-5876"],"issn-type":[{"value":"1479-5876","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,10,10]]},"assertion":[{"value":"29 March 2025","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"11 September 2025","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"10 October 2025","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Declarations"}},{"value":"Written informed consent was obtained from all participants in this study. The study protocol and informed consent form were approved by the Committee on Human Subject Research and Ethics of Yunnan University (2024100).","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Ethics approval and consent to participate"}},{"value":"Not applicable.","order":3,"name":"Ethics","group":{"name":"EthicsHeading","label":"Consent for publication"}},{"value":"All data generated or analyzed during this study are included in the manuscript and its additional information files.","order":4,"name":"Ethics","group":{"name":"EthicsHeading","label":"Availability of data and material"}},{"value":"Z.Z., C.L., T.Z. J.L. and X.Z. are the inventors of the patent (application number: 202410615729.3). The remaining authors declare no competing interests.","order":5,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"1077"}}