ELISA genotyping of hepatitis B virus in China with antibodies specific for genotypes B and C.
Diagnosis
Enzyme-linked immunosorbent assay (ELISA)
Genotype-specific antibody
Genotyping
Hepatitis B virus (HBV)
Journal
Scientific reports
ISSN: 2045-2322
Titre abrégé: Sci Rep
Pays: England
ID NLM: 101563288
Informations de publication
Date de publication:
12 Oct 2024
12 Oct 2024
Historique:
received:
13
05
2024
accepted:
09
10
2024
medline:
13
10
2024
pubmed:
13
10
2024
entrez:
12
10
2024
Statut:
epublish
Résumé
Hepatitis B virus (HBV) causes hepatitis B (HB) and distinct HBV genotypes can lead to different prognoses. However, HBV genotyping is rarely done in clinics, because the traditional method by PCR-based DNA sequencing is impractical for clinical diagnosis with tedious process and low success rate. Herein, we have established an ELISA-based genotyping method to quickly determine the HBV genotypes of HB patients in China. First, two commercial antibodies, 16D12 and 6H3 specific for HBV genotypes B and C respectively, are chosen as capture antibodies, since these two genotypes dominate in China. Then two home-made genotype-specific antibodies, B19 and C04, are used as the detection antibodies for genotypes B and C in sandwiched ELISA. The ELISA kit shows high sensitivity (> 95%) and specificity (> 95%) in detecting genotypes B and C of Chinese HB patients. Moreover, the ELISA kit has demonstrated higher success rate (98.7%) than PCR-based DNA sequencing (93.5%) and a commercial PCR-based genotyping kit (92.2%) for sera with HBV DNA ≥ 1000 IU/mL and HBsAg ≥ 250 IU/mL. Such an advantage is more obvious for the sera with HBV DNA < 1000 IU/mL. The kappa analysis between the ELISA and PCR-based DNA sequencing results exhibits a kappa of 0.836, indicating a good correlation.
Identifiants
pubmed: 39396069
doi: 10.1038/s41598-024-76023-7
pii: 10.1038/s41598-024-76023-7
doi:
Substances chimiques
DNA, Viral
0
Hepatitis B Antibodies
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
23884Subventions
Organisme : National Key Research and Development Program of China
ID : 2017ZX106080
Organisme : National Key Research and Development Program of China
ID : 2022YFE0138500
Organisme : National Key Research and Development Program of China
ID : 2017ZX106080
Organisme : National Key Research and Development Program of China
ID : 2017ZX106080
Organisme : National Key Research and Development Program of China
ID : 2017ZX106080
Organisme : National Key Research and Development Program of China
ID : 2017ZX106080
Informations de copyright
© 2024. The Author(s).
Références
Zheng, Q. et al. Single-cell landscape identifies the immunophenotypes and microenvironments of HBV-positive and HBV-negative liver cancer. Hepatol.Commun. 8, e0364. https://doi.org/10.1097/hc9.0000000000000364 (2024).
doi: 10.1097/hc9.0000000000000364
pubmed: 38251896
pmcid: 10805423
Kumar, M. et al. Asian Pacific association for the study of liver (APASL) guidelines: hepatitis B virus in pregnancy. Hepatol. Int. 16, 211–253 (2022).
doi: 10.1007/s12072-021-10285-5
pubmed: 35113359
Chen, L. et al. HBV core promoter mutations and AKT upregulate S-phase kinase-associated protein 2 to promote postoperative hepatocellular carcinoma progression. Sci. Rep. 6, 35917. https://doi.org/10.1038/srep35917 (2016).
doi: 10.1038/srep35917
pubmed: 27779207
pmcid: 5078796
Congly, S. E. et al. Characterization of hepatitis B virus genotypes and quantitative hepatitis B surface antigen titres in north American tertiary referral liver centres. Liver Int. 33, 1363–1369 (2013).
doi: 10.1111/liv.12222
pubmed: 23763288
Tong, S. & Revill, P. Overview of hepatitis B viral replication and genetic variability. J. Hepatol. 64, 4–16 (2016).
doi: 10.1016/j.jhep.2016.01.027
pubmed: 27084035
pmcid: 4834849
Kramvis, A. Genotypes and genetic variability of Hepatitis B Virus. Intervirology. 57, 141–150 (2014).
doi: 10.1159/000360947
pubmed: 25034481
Sun, B. et al. Origin and dispersal history of Hepatitis B virus in Eastern Eurasia. Nat. Commun. 15, 2951. https://doi.org/10.1038/s41467-024-47358-6 (2024).
doi: 10.1038/s41467-024-47358-6
pubmed: 38580660
pmcid: 10997587
Velkov, S., Ott, J. J., Protzer, U. & Michler, T. The Global Hepatitis B Virus Genotype Distribution Approximated from Available Genotyping Data. Genes. 9, 495. https://doi.org/10.3390/genes9100495 (2018).
Li, J. S. et al. Hepatitis B virus genotype A rarely circulates as an HBe-minus mutant: possible contribution of a single nucleotide in the precore region. J. Virol. 67, 5402–5410 (1993).
doi: 10.1128/jvi.67.9.5402-5410.1993
pubmed: 8350403
pmcid: 237941
Inoue, J. et al. Comparison of hepatitis B virus genotypes B and C among chronically hepatitis B virus-infected patients who received nucleos(t)ide analogs: a multicenter retrospective study. Hepatol. Res. 49, 1263–1274 (2019).
doi: 10.1111/hepr.13398
pubmed: 31254482
Bowden, D. S. & Thompson, A. J. New developments in HBV molecular diagnostics and quantitative serology. Hepatol. Int. 2, 3–11 (2008).
doi: 10.1007/s12072-008-9051-8
pubmed: 19669293
pmcid: 2716842
Sugauchi, F. et al. Epidemiologic and virologic characteristics of hepatitis B virus genotype B having the recombination with genotype C. Gastroenterology. 124, 925–932 (2003).
doi: 10.1053/gast.2003.50140
pubmed: 12671889
Locarnini, S., Littlejohn, M., Aziz, M. N. & Yuen, L. Possible origins and evolution of the hepatitis B virus (HBV). Semin Cancer Biol. 23, 561–575 (2013).
doi: 10.1016/j.semcancer.2013.08.006
pubmed: 24013024
Huy, T. T. T. et al. Characteristics of hepatitis B virus in Ghana: full length genome sequences indicate the endemicity of genotype E in West Africa. J. Med. Virol. 78, 178–184 (2006).
doi: 10.1002/jmv.20525
pubmed: 16372296
Arauz-Ruiz, P., Norder, H., Robertson, B. H. & Magnius, L. O. Genotype H: a new Amerindian genotype of hepatitis B virus revealed in Central America. J. Gen. Virol. 83, 2059–2073 (2002).
doi: 10.1099/0022-1317-83-8-2059
pubmed: 12124470
Ghany, M. G. et al. Characteristics of adults in the Hepatitis B Research Network in North America reflect their country of Origin and Hepatitis B Virus genotype. Clin. Gastroenterol. Hepatol. 13, 183–192 (2015).
doi: 10.1016/j.cgh.2014.06.028
pubmed: 25010003
Wang, Z. et al. A new intertype recombinant between genotypes C and D of hepatitis B virus identified in China. J. Gen. Virol. 86, 985–990 (2005).
doi: 10.1099/vir.0.80771-0
pubmed: 15784891
Tang, L. S. Y., Covert, E., Wilson, E. & Kottilil, S. Chronic Hepatitis B Infection: A Review. JAMA 319, 1802–1813 (2018).
doi: 10.1001/jama.2018.3795
pubmed: 29715359
Ito, K., Yoneda, M., Sakamoto, K. & Mizokami, M. Virological and clinical characteristics of Hepatitis B Virus genotype A. J. Gastroenterol. 53, 18–26 (2018).
doi: 10.1007/s00535-017-1367-5
pubmed: 28687901
Erhardt, A. et al. Response to interferon alfa is hepatitis B virus genotype dependent: genotype A is more sensitive to interferon than genotype D. Gut. 54, 1009–1013 (2005).
doi: 10.1136/gut.2004.060327
pubmed: 15951551
pmcid: 1774609
Shen, F. et al. Hepatitis B virus sensitivity to interferon-alpha in hepatocytes is more associated with cellular interferon response than with viral genotype. Hepatology. 67, 1237–1252 (2018).
doi: 10.1002/hep.29609
pubmed: 29059468
Deng, M., Tong, M., Fu, F. & Wei, D. Comparative untargeted metabolomics analysis of serum metabolic alterations in patients infected with hepatitis B virus genotypes B and C. Arab. J. Chem. 16, 105155. https://doi.org/10.1016/j.arabjc.2023.105155 (2023).
Chu, Y. J. et al. HBV genotype-dependent association of HLA variants with the serodecline of HBsAg in chronic hepatitis B patients. Sci. Rep. 13, 359. https://doi.org/10.1038/s41598-023-27570-y (2023).
doi: 10.1038/s41598-023-27570-y
pubmed: 36611047
pmcid: 9825396
Li, J. et al. Clinical isolates of hepatitis B virus genotype C have higher in vitro transmission efficiency than genotype B isolates. J. Med. Virol. 95, e28879. https://doi.org/10.1002/jmv.28879 (2023).
doi: 10.1002/jmv.28879
pubmed: 37314050
pmcid: 10404337
Lin, C. L. & Kao, J. H. Hepatitis B virus genotypes and variants. Cold Spring Harb Perspect. Med. 5, a021436. https://doi.org/10.1101/cshperspect.a021436 (2015).
doi: 10.1101/cshperspect.a021436
Chan, H. L. Y. et al. Hepatitis B Virus genotype C is Associated with more severe liver fibrosis than Genotype B. Clin. Gastroenterol. Hepatol. 7, 1361–1366 (2009).
doi: 10.1016/j.cgh.2009.08.004
pubmed: 19683072
Ni, Y. H. et al. Clinical relevance of hepatitis B virus genotype in children with chronic infection and hepatocellular carcinoma. Gastroenterology. 127, 1733–1738 (2004).
doi: 10.1053/j.gastro.2004.09.048
pubmed: 15578511
Watanabe, K. et al. Comparative study of genotype B and C hepatitis B virus-induced chronic hepatitis in relation to the basic core promoter and precore mutations. J. Gastroenterol. Hepatol. 20, 441–449 (2005).
doi: 10.1111/j.1440-1746.2004.03572.x
pubmed: 15740490
Revill, P. A. et al. The evolution and clinical impact of hepatitis B virus genome diversity. Nat. Rev. Gastroenterol. Hepatol. 17, 618–634 (2020).
doi: 10.1038/s41575-020-0296-6
pubmed: 32467580
Sukowati, C. H. C., Jayanti, S., Turyadi, T., Muljono, D. H. & Tiribelli, C. Hepatitis B virus genotypes in precision medicine of hepatitis B-related hepatocellular carcinoma: where we are now. World J. Gastrointest. Oncol. 16, 1097–1103 (2024).
doi: 10.4251/wjgo.v16.i4.1097
pubmed: 38660644
pmcid: 11037070
Lau, K. C., Osiowy, C. & Coffin, C. S. Hepatitis B virus (HBV) genome detection and genotyping in virally suppressed patients using nested polymerase chain reaction-based Sanger sequencing. Diagn. Microbiol. Infect. Dis. 93, 318–324 (2019).
doi: 10.1016/j.diagmicrobio.2018.10.015
pubmed: 30471842
Irshad, M., Gupta, P., Mankotia, D. S. & Ansari, M. A. Multiplex qPCR for serodetection and serotyping of hepatitis viruses: a brief review. World J. Gastroenterol. 20, 4824-4834 (2016).
Wang, W. et al. Establishment of a novel two-probe real-time PCR for simultaneously quantification of hepatitis B virus DNA and distinguishing genotype B from non-B genotypes. Clin. Chim. Acta. 437, 168–174 (2014).
doi: 10.1016/j.cca.2014.07.021
pubmed: 25066032
Chen, J. et al. Improved multiplex-PCR to identify hepatitis B virus genotypes A–F and subgenotypes B1, B2, C1 and C2. J. Clin. Virol. 38, 238–243 (2007).
doi: 10.1016/j.jcv.2006.12.007
pubmed: 17224304
Welzel, T. M. et al. Real-time PCR assay for detection and quantification of Hepatitis B virus genotypes a to G. J. Clin. Microbiol. 44, 3325–3333 (2006).
doi: 10.1128/jcm.00024-06
pubmed: 16954268
pmcid: 1594718
Liu, K. et al. Identifying the genotypes of Hepatitis B Virus (HBV) with DNA origami label. Small. 14, 1701718. https://doi.org/10.1002/smll.201701718 (2018).
Usuda, S. et al. Serological detection of hepatitis B virus genotypes by ELISA with monoclonal antibodies to type-specific epitopes in the preS2-region product. J. Virol. Methods. 80, 97–112 (1999).
doi: 10.1016/S0166-0934(99)00039-7
pubmed: 10403681
Bezuglova, L. V. et al. Trial results for ELISA test kits for HBsAg Subtype and Hepatitis B Virus genotype identification in human blood plasma. Mol. Genet. Microbiol.Virol. 35, 229–236 (2020).
doi: 10.3103/S0891416820040047
Zhang, R. et al. Determination of hepatitis B virus genotype by flow-through reverse dot blot. J. Clin. Virol. 39, 94–100 (2007).
doi: 10.1016/j.jcv.2007.01.014
pubmed: 17466578
Song, L. W. et al. Rapid fluorescent lateral-flow immunoassay for hepatitis B virus genotyping. Anal. Chem. 87, 5173–5180 (2015).
doi: 10.1021/ac504832c
pubmed: 25892477
Qiu, X. et al. A fast and low-cost genotyping method for hepatitis B virus based on pattern recognition in point-of-care settings. Sci. Rep. 6, 28274. https://doi.org/10.1038/srep28274 (2016).
doi: 10.1038/srep28274
pubmed: 27306485
pmcid: 4910285
Ohishi, W. & Chayama, K. Treatment of chronic hepatitis B with nucleos(t)ide analogues. Hepatol. Res. 42, 219–225 (2012).
doi: 10.1111/j.1872-034X.2011.00921.x
pubmed: 22176139
Ye, X. et al. Efficient inhibition of Hepatitis B Virus infection by a preS1-binding peptide. Sci. Rep. 6, 29391. https://doi.org/10.1038/srep29391 (2016).
doi: 10.1038/srep29391
pubmed: 27384014
pmcid: 4935942
Seeger, C. & Mason, W. S. Hepatitis B Virus Biology. Microbiol. Mol. Biol. Rev. 64, 51–68 (2000).
doi: 10.1128/MMBR.64.1.51-68.2000
pubmed: 10704474
pmcid: 98986
Meier, A., Mehrle, S., Weiss, T. S., Mier, W. & Urban, S. Myristoylated PreS1-domain of the hepatitis B virus L-protein mediates specific binding to differentiated hepatocytes. Hepatology. 58, 31–42 (2013).
doi: 10.1002/hep.26181
pubmed: 23213046
Hellstrom, U., Lindh, M., Krogsgaard, K. & Sylvan, S. Demonstration of an association between detection of IgG antibody reactivity towards the C-terminal region of the preS1 protein of hepatitis B virus and the capacity to respond to interferon therapy in chronic hepatitis B. J. Gastroenterol. Hepatol. 23, 804–810 (2008).
doi: 10.1111/j.1440-1746.2007.05174.x
pubmed: 17931371
Wai, C. T., Chu, C. J., Hussain, M. & Lok, A. S. F. HBV genotype B is associated with better response to interferon therapy in HBeAg(+) chronic hepatitis than genotype C. Hepatology. 36, 1425–1430 (2002).
doi: 10.1002/hep.1840360619
pubmed: 12447868
Kirkwood, T. B. L. & Tydeman, M. S. Design and analysis of accelerated degradation tests for the stability of biological standards II. A flexible computer program for data analysis. J. Biol. Stand. 12, 207–214 (1984).
doi: 10.1016/S0092-1157(84)80055-4
pubmed: 6547442
Lago, B. V. et al. Genetic variability of hepatitis B virus in acute and in different phases of chronic infection in Brazil. Sci. Rep. 14, 10742. https://doi.org/10.1038/s41598-024-60900-2 (2024).
doi: 10.1038/s41598-024-60900-2
pubmed: 38730249
pmcid: 11087654
Razavi-Shearer, D. et al. Global prevalence, treatment, and prevention of hepatitis B virus infection in 2016: a modelling study. Lancet Gastroenterol. Hepatol. 3, 383–403 (2018).
doi: 10.1016/S2468-1253(18)30056-6
Chiu, S. M. et al. Associations of HBV genotype B vs C infection with Relapse after Cessation of Entecavir or Tenofovir Therapy. Clin. Gastroenterol. Hepatol. 18, 2989–2997 (2020).
doi: 10.1016/j.cgh.2020.04.048
pubmed: 32353534
Peng, X., Chen, L. & Xiao, M. Entecavir/peginterferon alfa-2a combination in the treatment of two genotypes of chronic hepatitis B patients with lamivudine resistance. Trop. J. Pharm. Res. 22, 901–907 (2023).
doi: 10.4314/tjpr.v22i4.25
Zhang, B. et al. Hepatitis B virus genotype is an independent prognostic factor of telbivudine and tenofovir treatment in hepatitis B surface antigen-positive pregnant women. Food Sci. Nutr. 10, 3–11 (2022).
doi: 10.1002/fsn3.2619
pubmed: 35035905
Jardi, R. et al. Analysis of hepatitis B genotype changes in chronic hepatitis B infection: influence of antiviral therapy. J. Hepatol. 49, 695–701 (2008).
doi: 10.1016/j.jhep.2008.03.030
pubmed: 18490074
Wen, J. et al. Hepatitis B virus genotype, mutations, human leukocyte antigen polymorphisms and their interactions in hepatocellular carcinoma: a multi-centre case-control study. Sci. Rep. 5, 16489. https://doi.org/10.1038/srep16489 (2015).
doi: 10.1038/srep16489
pubmed: 26568165
pmcid: 4644975
Varghese, N. et al. Review of related factors for persistent risk of Hepatitis B Virus-Associated Hepatocellular Carcinoma. Cancers. 16, 777. https://doi.org/10.3390/cancers16040777 (2024).
Wiegand, S. B. et al. Soluble immune markers in the different phases of chronic hepatitis B virus infection. Sci. Rep. 9, 14118. https://doi.org/10.1038/s41598-019-50729-5 (2019).
doi: 10.1038/s41598-019-50729-5
pubmed: 31575964
pmcid: 6773856
Utama, A. et al. Low prevalence of hepatitis B virus pre-s deletion mutation in Indonesia. J. Med. Virol. 83, 1717–1726 (2011).
doi: 10.1002/jmv.22172
pubmed: 21837787
Gao, Y. et al. Serum Hepatitis B Virus DNA, RNA, and HBsAg: which correlated better with Intrahepatic Covalently closed circular DNA before and after Nucleos(t)ide Analogue Treatment? J. Clin. Microbiol. 55, 2972–2982 (2017).
doi: 10.1128/JCM.00760-17
pubmed: 28747369
pmcid: 5625383
Robert, X. & Gouet, P. Deciphering key features in protein structures with the new ENDscript server. Nucleic Acids Res. 42, W320–W324 (2014).
doi: 10.1093/nar/gku316
pubmed: 24753421
pmcid: 4086106