Significance of Hepatitis B Virus Diagnosis by Real-time Polymerase Chain Reaction over Serological Markers in Hepatitis B Virus Patients
Abstract
Hepatitis B virus (HBV) is the leading cause of viral hepatitis, as currently over 2 billion people have HBV infection worldwide. Nucleic acid assay and quantitative hepatitis B surface antigen (HBsAg) have been developed for diagnostic and therapeutic monitoring of patients with HBV infection. These tests might also show correlation between HBV DNA and HBs serostatus. The study aimed to find and analyze the frequency and impact of HBsAg seropositivity among patients revealed HBV DNA negative level through quantitative estimation of both seromarkers. Real-time polymerase chain reaction (RT-PCR) and Elecsys assays were used for quantitative estimation of HBV DNA and HBs antigen, respectively. A total of 256 blood samples were used from patients referred for either diagnostic purpose and/or HBV viral load monitoring after antiviral therapy. Blood profile analysis showed 12.26% HBs antigen seropositivity among patients revealed negative for nucleic acid assay for HBV DNA. Positive HBs antigen titers ranged from 1000–50,000 COI, with seronegative anti-HBs antibody test for all samples tested positive for HBs antigen. This study delineated that negative or undetectable quantitation of HBV DNA level does not exclude HBV infection; as the level might fluctuate in different phases of HBV replication. This gives an impression and raising a question about significance of replacing test for HBsAg with quantitation of HBV DNA PCR assay. Thus, the study refers to a special HBV profile outside the classical pattern.
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References
World Health Organization. “World Health Organization Hepatitis B Factsheet”. Geneva: World Health Organization, 2017.
S. Baig, A. A. Siddiqui, W. Ahmed, H. Qureshi and A. Arif. “The association of complex liver disorders with HBV genotypes prevalent in Pakistan”. Virology Journal, vol. 4, no. 1, p. 128, 2007.
G. Fattovich. “Natural history of hepatitis B”. Journal of Hepatology, vol. 39, pp. 50-58, 2003.
I. Pita, A. M. Horta-Vale, H. Cardoso and G. Macedo. “Hepatitis B inactive carriers: An overlooked population?” GE Portuguese Journal of Gastroenterology, vol. 21, no. 6, pp. 241-249, 2014.
L. M. Villar, H. M. Cruz, J. R. Barbosa, C. S. Bezerra, M. M. Portilho and L. de Paula Scalioni. “Update on hepatitis B and C virus diagnosis”. World Journal of Virology, vol. 4, no. 4, p. 323, 2015.
I. J. Radii and A. M. Saud. “Comparative assessment between serological and molecular diagnosis for patients groups with hepatitis b virus”. International Journal of Current Microbiology and Applied Science, vol. 6, no. 3, pp. 742-748, 2017.
T. C. Tseng, C. J. Liu, H. C. Yang, T. H. Su, C. C. Wang, C. L. Chen and J. H. Kao. “High levels of hepatitis B surface antigen increase risk of hepatocellular carcinoma in patients with low HBV load”. Gastroenterology, vol. 142, no. 5, pp. 1140-1149, 2012.
M. Kawanaka, K. Nishino, J. Nakamura, T. Oka, N. Urata, D. Goto and G. Yamada. “Quantitative levels of hepatitis B virus DNA and surface antigen and the risk of hepatocellular carcinoma in patients with hepatitis B receiving long-term nucleos (t) ide analogue therapy”. Liver Cancer, vol. 3, no. 1, pp. 41-52, 2014.
T. C. Tseng, C. J. Liu, T. H. Su, C. C. Wang, C. L. Chen, P. J. Chen and J. H. Kao. “Serum hepatitis B surface antigen levels predict surface antigen loss in hepatitis B e antigen seroconverts”. Gastroenterology, vol. 141, no. 2, pp. 517-525, 2011.
K. M. Weinberger, E. Wiedenmann, S. Böhm and W. Jilg. “Sensitive and accurate quantitation of hepatitis B virus DNA using a kinetic luorescence detection system (TaqMan PCR)”. Journal of Virological Methods, vol. 85, no. 1-2, pp. 75-82, 2000.
H. J. Lee, S. Y. Kim, S. M. Lee, J. Heo, H. H. Kim, C. L. Chang and H. C. Son. “Elecsys hepatitis B surface antigen quantitative assay: Performance evaluation and correlation with hepatitis B virus DNA during 96 weeks of follow-up in chronic hepatitis B patients”. Annals of Laboratory Medicine, vol. 32, no. 6, pp. 420-425, 2012.
A. S. Lok and B. J. McMahon. “Chronic hepatitis B: Update 2009”. Hepatology, vol. 50, no. 3, pp. 661-662, 2009.
European Association for the Study of the Liver. “EASL clinical practice guidelines: Management of chronic hepatitis B virus infection”. Journal of Hepatology, vol. 57, no. 1, pp. 167-185, 2012.
Y. F. Liaw, J. H. Kao, T. Piratvisuth, H. L. Y. Chan, R. N. Chien, C. J. Liu and D. Amarapurkar. “Asian-Pacific consensus statement on the management of chronic hepatitis B: A 2012 update”. Hepatology International, vol. 6, no. 3, pp. 531-561, 2012.
H. Lik-Yuen Chan, G. Lai-Hung Wong, C. H. Tse, H. Y. Chan and V. Wai-Sun Wong. “Viral determinants of hepatitis b surface antigen seroclearance in hepatitis b e antigen-negative chronic hepatitis b patients”. Journal of Infectious Diseases, vol. 204, no. 3, pp. 408-414, 2011.
J. Cheng, C. Sun, Y. Chen, Y. Dai, X. U. Zhiliang, G. Sun and L. I. Xiaojun. “Molecular analysis on chronic hepatitis B patients with low-level HBsAg”. Chinese Journal of Laboratory Medicine, vol. 32, no. 10, pp. 1128-1132, 2009.
J. Cheng, C. Sun, Y. Chen, Z. Xu, G. Wang, G. Sun and X. Li. “Analysis on clinical features and immunity in chronic hepatitis B virus infected patients with low-level HBsAg”. African Journal of Microbiology Research, vol. 4, no. 7, pp. 547-550, 2010.
A. Jeffery-Smith, J. Hubb, A. Oliver and C. Y. Tong. “An apparent low level of hepatitis B surface antigen (HBsAg) in the presence of significant viral replication”. Journal of Clinical Virology: The Official Publication of the Pan American Society for Clinical Virology, vol. 77, p. 111, 2016.
T. Nguyen, A. J. Thompson, S. Bowden, C. Croagh, S. Bell, P. V Desmond and S. A. Locarnini. “Hepatitis B surface antigen levels during the natural history of chronic hepatitis B: A perspective on Asia”. Journal of Hepatology, vol. 52, no. 4, pp. 508-513, 2010.
S. T. Balaky, S. G. Hussain, A. A. Khaleel, F. T. Sabeer and A. H. Mawlood. “Molecular diagnosis in differentiating active and inactive forms of
hepatitis b virus carriers”. Journal of University of Babylon for Pure and Applied Sciences, vol. 26, no. 10, pp. 41-45, 2018.
M. C. Kuhns, S. H. Kleinman, A. L. McNamara, B. Rawal, S. Glynn and M. P. Busch. “Lack of correlation between HBsAg and HBV DNA levels in blood donors who test positive for HBsAg and anti-HBc: Implications for future HBV screening policy”. Transfusion, vol. 44, no. 9, pp. 1332-1339, 2004.
A. S. K. Al-Suraifi, A. D. J. Al-Rubaie, S. M. G. Al-Mayahie and N. M. M. Al-Abedy. “Unusual HBV mixed genotype infections among hepatitis type B Iraqi patients in Wasit Province/Iraq”. International Journal of Biomedical Engineering and Clinical Science, vol. 2, no. 1, pp. 1-7, 2016.
I. A. A. Khaled, O. M. Mahmoud, A. F. Saleh and E. A. Baioumi. “Prevalence of HBV genotypes in Egypt among hepatitis patients”. Journal of American Science, vol. 6, no. 11, pp. 185-190, 2010.
W. K. Roth, M. Weber, D. Petersen, C. Drosten, S. Buhr, W. Sireis and E. Seifried. “NAT for HBV and anti-HBc testing increase blood safety”. Transfusion, vol. 42, no. 7, pp. 869-875, 2002.
S. Sato, W. Ohhashi, H. Ihara, S. Sakaya, T. Kato and H. Ikeda. “Comparison of the sensitivity of NAT using pooled donor samples for HBV and that of a serologic HBsAg assay”. Transfusion, vol. 41, no. 9, pp. 1107-1113, 2001.
H. Hennig, I. Puchta, J. Luhm, P. Schlenke, S. Goerg and H. Kirchner. “Frequency and load of hepatitis B virus DNA in first time blood donors with antibodies to hepatitis B core antigen”. Blood, vol. 100, no. 7, pp. 2637-2641, 2002.
M. C. Kuhns, A. L. McNamara and R. Perrillo. “Intermittent PCR positivity for hepatitis B viral DNA in surface antigen negative anti-core positive individuals”. Transfusion, vol. 39, no. 10, pp. 95S-95S, 1999.
Y. J. Kim, H. C. Cho, M. S. Choi, J. H. Lee, K. C. Koh, B. C. Yoo and S. W. Paik. “The change of the quantitative HBsAg level during the natural course of chronic hepatitis B”. Liver International, vol. 31, no. 6, pp. 817-823, 2011.
J. Jaroszewicz, B. C. Serrano, K. Wursthorn, K. Deterding, J. Schlue, R. Raupach and M. Cornberg. “Hepatitis B surface antigen (HBsAg) levels in the natural history of hepatitis B virus (HBV)-infection: A European perspective”. Journal of Hepatology, vol. 52, no. 4, pp. 514-522, 2010.
A. Alghamdi, N. Aref, M. El-Hazmi, W. Al-Hamoudi, K. Alswat, A. Helmy and A. A. Abdo. “Correlation between hepatitis B surface antigen titers and HBV DNA levels”. Saudi Journal of Gastroenterology: Official Journal of the Saudi Gastroenterology Association, vol. 19, no. 6, p. 252, 2013.
P. P. Primadharsini and I. Wibawa. “Correlation between quantitative HBsAg and HBV-DNA in chronic hepatitis B infection”. Indonesian Journal of Gastroenterology, Hepatology, and Digestive Endoscopy, vol. 14, no. 1, pp. 9-12, 2013.
M. Z. Hong, W. Q. Huang, F. Min, J. C. Xu, Z. Lin, K. N. Fang and J. S. Pan. “Enhanced HBsAg synthesis correlates with increased severity of fibrosis in chronic hepatitis B patients”. PLoS One, vol. 9, no. 1, p. e87344, 2014.
O. Günal, S. Barut, I. Etikan, F. Duygu, U. Tuncel and M. Sünbül. “Relation between serum quantitative HBsAg, ALT and HBV DNA levels in HBeAg negative chronic HBV infection”. Turkish Journal of Gastroenterology, vol. 25, no. 1, pp. 142-146, 2014.
G. Li, J. Wang, Y. Bao, L. Zheng, K. Ge, X. Zhou and G. Chen. “Clinical significance of serum HBsAg levels, HBsAg/HBV DNA ratio, and association with liver inflammation activity in HBeAg-positive chronic hepatitis B”. Zhonghua Gan Zang Bing Za Zhi=Zhonghua Ganzangbing Zazhi=Chinese Journal of Hepatology, vol. 23, no. 1, pp. 40-45, 2015.
A. Ganji, A. Esmaeilzadeh, K. Ghafarzadegan, H. Helalat, H. Rafatpanah and A. Mokhtarifar. “Correlation between HBsAg quantitative assay results and HBV DNA levels in chronic HBV”. Hepatitis Monthly, vol. 11, no. 5, p. 342, 2011.
R. M. Mukherjee, J. Arava, P. B. Reddy, P. N. Rao, R. Gupta and D. N. Reddy. “Role of hepatitis b virus surface antigen quantification in E antigen negative chronic hepatitis B infection”. Global Journal of Science Frontier Research, vol. 13, no. 1, pp. 1-5, 2013.
H. Park, J. M. Lee, J. H. Seo, H. S. Kim, S. H. Ahn, D. Y. Kim and J. Y. Park. “Predictive value of HBsAg quantification for determining the clinical course of genotype C HBeAg-negative carriers”. Liver International, vol. 32, no. 5, pp. 796-802, 2012.
A. J. Thompson, T. Nguyen, D. Iser, A. Ayres, K. Jackson, M. Littlejohn and G. K. Lau. “Serum hepatitis B surface antigen and hepatitis B e antigen titers: Disease phase influences correlation with viral load and intrahepatic hepatitis B virus markers”. Hepatology, vol. 51, no. 6, pp. 1933-1944, 2010.
M. Martinot-Peignoux, M. Lapalus, C. Laouénan, O. Lada, A. C. F. Netto-Cardoso, N. Boyer and P. Marcellin. “Prediction of disease reactivation in asymptomatic hepatitis B e antigennegative chronic hepatitis B patients using baseline serum measurements of HBsAg and HBV-DNA”. Journal of Clinical Virology, vol. 58, no. 2, pp. 401-407, 2013.
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