Prevalence and Characteristics of Precore Mutation in Iran and Its Correlation with Genotypes of Hepatitis B
Keywords:
Precore mutants; Genotypes; Hepatitis B; RFLPAbstract
Introduction: Mutation of the HBV precore gene prevents the production of HBeAg, which is an important target for immune responses. Distribution of this mutation varies along with frequency of HBV genotypes in accordance with geographic and ethnic variations. The general objective of this study was to evaluate the prevalence and characteristics of precore mutation in Iran and its correlation with genotypes of hepatitis B.
Methods: In this cross-sectional study, viral DNA of 182 Iranian hepatitis B surface antigen positive patients who were admitted to Bandar Abbas Blood Transfusion Organization in 2012 and 2013 was retrieved from their serum samples. HBeAg, anti-HBe, and anti-HBc IgM diagnostic tests were performed using ELISA kits. Precore and Pre-S regions were amplified using specific primers and PCR thereafter to determine the genotypes; precore mutation, PCR, and restriction fragment length polymorphism (RFLP) methods also were applied. SPSS version 12 was used for data analysis by Mann–Whitney U test, Fisher’s exact probability test, and t-test.
Results: A total of 62 patients (34.1%) had precore mutation (A1896G), and genotype D was the predominant genotype in these patients, which was followed by an unknown genotype that was suspected for genotype B. Interestingly, the relationships between precore mutation and HBeAg (p=0.037) and genotype D (p=0.005) were significant; however, no correlation was observed between this mutation and acute or chronic hepatitis and sex of patients.
Conclusion: This study found high prevalence of precore mutations in southern Iran, which was significantly associated with HBeAg and genotype D.
References
Takahashi K, Aoyama K, Ohno N, Iwata K, Akahane Y, Baba K, et al. The precore/core promoter mutant
(T1762A1764) of hepatitis B virus: clinical significance and an easy method for detection. J Gen Virol.
; 76 ( Pt 12): 3159-64. doi: 10.1099/0022-1317-76-12-3159. PMID: 8847524.
Lee WM. Hepatitis B virus infection. N Engl J Med. 1997; 337(24): 1733-45. doi:
1056/NEJM199712113372406. PMID: 9392700.
Garcia PD, Ou JH, Rutter WJ, Walter P. Targeting of the hepatitis B virus precore protein to the
endoplasmic reticulum membrane: after signal peptide cleavage translocation can be aborted and the
product released into the cytoplasm. J Cell Biol. 1988; 106(4): 1093-104. doi: 10.1083/jcb.106.4.1093.
PMID: 3283145, PMCID: PMC2114996.
Chen IH, Huang CJ, Ting LP. Overlapping initiator and TATA box functions in the basal core promoter of
hepatitis B virus. J Virol. 1995; 69(6): 3647-57. PMID: 7745713, PMCID: PMC189080.
Chen HS, Kew MC, Hornbuckle WE, Tennant BC, Cote PJ, Gerin JL, et al. The precore gene of the
woodchuck hepatitis virus genome is not essential for viral replication in the natural host. J Virol. 1992;
(9): 5682-4. PMID: 1501300, PMCID: PMC289138.
Chisari FV, Ferrari C. Hepatitis B virus immunopathogenesis. Annu Rev Immunol. 1995; 13: 29-60. doi:
1146/annurev.iy.13.040195.000333. PMID: 7612225.
Akahane Y, Yamanaka T, Suzuki H, Sugai Y, Tsuda F, Yotsumoto S, et al. Chronic active hepatitis with
hepatitis B virus DNA and antibody against e antigen in the serum: Disturbed synthesis and secretion of e
antigen from hepatocytes due to a point mutation in the precore region. Gastroenterology. 1990; 99(4):
-9. doi: 10.1016/0016-5085(90)90632-B. PMID: 2394332.
Okada K, Kamiyama I, Inomata M, Imai M, Miyakawa Y, Mayumi M. e antigen and anti-e in the serum of
asymptomatic carrier mothers as indicators of positive and negative transmission of hepatitis B virus to
their infants. N Eng J Med. 1976; 294(14): 746-9. doi: 10.1056/NEJM197604012941402. PMID: 943694.
Carman WF, Jacyna MR, Hadziyannis S, Karayiannis P, McGarvey MJ, Makris A, et al. Mutation
preventing formation of hepatitis B e antigen in patients with chronic hepatitis B infection. Lancet. 1989;
(8663): 588-91. doi: 10.1016/S0140-6736(89)90713-7. PMID: 2570285.
Brunetto MR, Giarin MM, Oliveri F, Chiaberge E, Baldi M, Alfarano A, et al. Wild-type and e antigen- minus hepatitis B viruses and course of chronic hepatitis. Proc Natl Acad Sci U S A. 1991; 88(10): 4186- 90. doi: 10.1073/pnas.88.10.4186. PMID: 2034663, PMCID: PMC51623.
Brunetto M. Identification of HBV variants which cannot produce pre-core derived HBeAg and may be
responsible for severe hepatitis. Ital J Gastroenterol. 1989; 21: 151-4. Available from:
http://ci.nii.ac.jp/naid/10006179330/.
Zuckerman AJ, Zuckerman JN. Molecular epidemiology of hepatitis B virus mutants. J med virol. 1999;
(3): 193-5. doi: 10.1002/(SICI)1096-9071(199907)58:3<193::AID-JMV1>3.0.CO;2-X. PMID:
López JL, Mbayed VA, Telenta PF, González JE, Campos RH. ‘Hbe minus’ mutants of hepatitis B virus.
Molecular characterization and its relation to viral genotypes. Virus Res. 2002; 87(1): 41-9. doi:
1016/S0168-1702(02)00078-3. PMID: 12135788.
Günther S, Meisel H, Reip A, Miska S, Krüger DH, Will H. Frequent and rapid emergence of mutated pre- C sequences in HBV from e-antigen positive carriers who seroconvert to anti-HBe during interferon
treatment. Virology. 1992; 187(1): 271-9. doi: 10.1016/0042-6822(92)90315-G. PMID: 1736529.
Sterneck M, Will H. Naturally occurring variants of hepatitis B virus. Adv Virus Res. 1999; 52: 25. doi:
1016/S0065-3527(08)60298-5.
Lindh M, Hannoun C, Dhillon AP, Norkrans G, Horal P. Core promoter mutations and genotypes in
relation to viral replication and liver damage in East Asian hepatitis B virus carriers. J Infect Dis. 1999;
(4): 775-82. doi: 10.1086/314688. PMID: 10068571.
Stuyver L, De Gendt S, Van Geyt C, Zoulim F, Fried M, Schinazi RF, et al. A new genotype of hepatitis B
virus: complete genome and phylogenetic relatedness. J Gen Virol. 2000; 81(Pt 1): 67-74. doi:
1099/0022-1317-81-1-67. PMID: 10640543.
Arauz-Ruiz P, Norder H, Robertson BH, Magnius LO. Genotype H: a new Amerindian genotype of
hepatitis B virus revealed in Central America. J Gen Virol. 2002; 83(8): 2059-73. doi: 10.1099/0022-1317- 83-8-2059. PMID: 12124470.
Magnius LO, Norder H. Subtypes, genotypes and molecular epidemiology of the hepatitis B virus as
reflected by sequence variability of the S-gene. Intervirology. 1995; 38(1-2): 24-34. doi:
1159/000150411. PMID: 8666521.
Okamoto H, Tsuda F, Sakugawa H, Sastrosoewignjo RI, Imai M, Miyakawa Y, et al. Typing hepatitis B
virus by homology in nucleotide sequence: comparison of surface antigen subtypes. J Gen Virol. 1988;
(Pt 10): 2575-83. doi: 10.1099/0022-1317-69-10-2575. PMID: 3171552.
Norder H, Hammas B, Löfdahl S, Couroucé AM, Magnius LO. Comparison of the amino acid sequences of
nine different serotypes of hepatitis B surface antigen and genomic classification of the corresponding
hepatitis B virus strains. J Gen Virol. 1992; 73(Pt 5): 1201-8. doi: 10.1099/0022-1317-73-5-1201. PMID:
Norder H, Couroucé AM, Magnius LO. Complete genomes, phylogenetic relatedness, and structural
proteins of six strains of the hepatitis B virus, four of which represent two new genotypes. Virology. 1994;
(2): 489-503. doi: 10.1006/viro.1994.1060. PMID: 8291231.
Guo W, Chen M, Yen TS, Ou JH. Hepatocyte-specific expression of the hepatitis B virus core promoter
depends on both positive and negative regulation. Mol Cell Biol. 1993; 13(1): 443-8. doi:
1128/MCB.13.1.443. PMID: 8417343, PMCID: PMC358924.
Guo WT, Bell KD, Ou JH. Characterization of the hepatitis B virus EnhI enhancer and X promoter
complex. J Virol. 1991; 65(12): 6686-92. PMID: 1658368. PMCID: PMC250743.
Norder H, Hammas B, Lee SD, Bile K, Couroucé AM, Mushahwar IK, et al. Genetic relatedness of
hepatitis B viral strains of diverse geographical origin and natural variations in the primary structure of the
surface antigen. J Gen Virol. 1993; (Pt 7): 1341-8. doi: 10.1099/0022-1317-74-7-1341. PMID: 8336122.
Orito E, Ichida T, Sakugawa H, Sata M, Horiike N, Hino K, et al. Geographic distribution of hepatitis B
virus (HBV) genotype in patients with chronic HBV infection in Japan. Hepatology. 2001; 34(3): 590-4.
doi: 10.1053/jhep.2001.27221. PMID: 11526547.
Ashouri FP, Rasekhi S. A Review on Medical Students Mental Health Problems and Proposed Solutions.
International Electronic Journal of Medicine. 2015; 4(1): 23-31.
Ashouri FP, Rasekhi S. Correlation between Religious Beliefs with Mental Health and Academic
Performance in Medical Students. International Electronic Journal of Medicine. 2016; 5(1): 1-6.
Pivert A, Servant‐Delmas A, Lunel‐Fabiani F, Guillou‐Guillemette L, Laperche S, Ducancelle A.
Correlation between the promoter basal core and precore mutations and HBsAg quantification in French
blood donors infected with hepatitis B virus. J Med Virol. 2015; 87(3): 529-35. doi: 10.1002/jmv.24064.
PMID: 25170961.
Shiina S, Fujino H, Uta Y, Tagawa K, Unuma T, Yoneyama M, et al. Relationship of HBsAg subtypes with
HBeAg/anti-HBe status and chronic liver disease. Part I: Analysis of 1744 HBsAg carriers. Am J
Gastroenterol. 1991; 86(7): 866-71. PMID: 2058629.
Kidd-Ljunggren K, Miyakawa Y, Kidd AH. Genetic variability in hepatitis B viruses. J Gen Virol. 2002;
(Pt 6): 1267-80. doi: 10.1099/0022-1317-83-6-1267. PMID: 12029141.
Hadziyannis SJ, Vassilopoulos D. Hepatitis B e antigen–negative chronic hepatitis B. Hepatology. 2001;
(4 Pt 1): 617-24. doi: 10.1053/jhep.2001.27834. PMID: 11584355.
Grandjacques C, Pradat P, Stuyver L, Chevallier M, Chevallier P, Pichoud C, et al. Rapid detection of
genotypes and mutations in the pre-core promoter and the pre-core region of hepatitis B virus genome:
correlation with viral persistence and disease severity. J Hepatol. 2000; 33(3): 430-9. doi: 10.1016/S0168- 8278(00)80279-2. PMID: 11019999.
Lindh M, Furuta Y, Vahlne A, Norkrans G, Horal P. Emergence of precore TAG mutation during hepatitis
B e seroconversion and its dependence on pregenomic base pairing between nucleotides 1858 and 1896. J
Infect Dis. 1995; 172(5): 1343-7. doi: 10.1093/infdis/172.5.1343. PMID: 7594674.
Ayed K, Gorgi Y, Ayed-Jendoubi S, Aouadi H, Sfar I, Najjar T, et al. Hepatitis B virus genotypes and
precore/core-promoter mutations in Tunisian patients with chronic hepatitis B virus infection. J Infect.
; 54(3): 291-7. doi: 10.1016/j.jinf.2006.05.013. PMID: 16911832.
Lok AS, Akarca U, Greene S. Mutations in the pre-core region of hepatitis B virus serve to enhance the
stability of the secondary structure of the pre-genome encapsidation signal. Proc Natl Acad Sci U S A.
; 91(9): 4077-81. PMID: 8171038, PMCID: PMC43726.
Orito E, Mizokami M, Sakugawa H, Michitaka K, Ishikawa K, Ichida T, et al. A case-control study for
clinical and molecular biological differences between hepatitis B viruses of genotypes B and C. Japan HBV
Genotype Research Group. Hepatology. 2001; 33(1): 218-23. doi: 10.1053/jhep.2001.20532. PMID:
Rodriguez‐Frias F, Buti M, Jardi R, Cotrina M, Viladomiu L, Esteban R, et al. Hepatitis B virus infection:
precore mutants and its relation to viral genotypes and core mutations. Hepatology. 1995; 22(6): 1641-7.
doi: 10.1002/hep.1840220605. PMID: 7489968.
Li JS, Tong SP, Wen YM, Vitvitski L, Zhang Q, Trépo C. Hepatitis B virus genotype A rarely circulates as
an HBe-minus mutant: possible contribution of a single nucleotide in the precore region. J Virol. 1993;
(9): 5402-10. PMID: 8350403, PMCID: PMC237941.
Meldal BH, Moula NM, Barnes IH, Boukef K, Allain JP. A novel hepatitis B virus subgenotype, D7, in
Tunisian blood donors. J Gen Virol. 2009; 90(Pt 7): 1622-8. doi: 10.1099/vir.0.009738-0. PMID:
Bahri O, Cheikh I, Hajji N, Djebbi A, Maamouri N, Sadraoui A, et al. Hepatitis B genotypes, precore and
core promoter mutants circulating in Tunisia. J Med Virol. 2006; 78(3): 353-7. doi: 10.1002/jmv.20554.
PMID: 16419125.
Borchani-Chabchoub I, Gargouri A, Mokdad-Gargouri R. Genotyping of Tunisian hepatitis B virus isolates
based on the sequencing of preS2 and S regions. Microbes Infect. 2000; 2(6): 607-12. doi: 10.1016/S1286- 4579(00)00365-8. PMID: 10884611.
Bodhiphala P, Chaturachumroenchai S, Chiewsilp P, Pruksananonda P. Detection of HBV genome by gene
amplification method in HBsAg negative blood donors. J Med Assoc Thai. 1999; 82(5): 491-5. PMID:
Alam-mehrjerdi Z, Mokri A, Dolan K. Methamphetamine use and treatment in Iran: a systematic review
from the most populated Persian Gulf country. Asian J Psychiatr. 2015; 16: 17-25. doi:
1016/j.ajp.2015.05.036. PMID: 26123235.
Hamadiyan H, Pour Ashouri F, Rasekhi S. Growth of Head Circumference, Weight and Height from Birth
to 18 Months in the South of Iran. International Electronic Journal of Medicine. 2015; 4(1): 1-5.
Yadollahi S, Hamadiyan H, Pour Ashouri F, Fallahi S, Parvizpanah A, Rasekhi S. Trends in the Lipid
Profile, Mean Age and Fatality of Patients with Myocardial Infarction in the South of Iran from 2008 to
Int J Med Res Health Sci. 2016; 5(6): 221-8.
Lindh M, Gonzalez JE, Norkrans G, Horal P. Genotyping of hepatitis B virus by restriction pattern analysis
of a pre-S amplicon. J Virol Methods. 1998; 72(2): 163-74. doi: 10.1016/S0166-0934(98)00026-3. PMID:
Okamoto H, Yotsumoto S, Akahane Y, Yamanaka T, Miyazaki Y, Sugai Y, et al. Hepatitis B viruses with
precore region defects prevail in persistently infected hosts along with seroconversion to the antibody
against e antigen. J Virol. 1990; 64(3): 1298-303. PMID: 2304145, PMCID: PMC249247.
Rezende RE, Fonseca BA, Ramalho LN, Zucoloto S, Pinho JR, Bertolini DA, et al. The precore mutation is
associated with severity of liver damage in Brazilian patients with chronic hepatitis B. J Clin Virol. 2005;
(1): 53-9. doi: 10.1016/j.jcv.2004.08.001. PMID: 15572007.
Vivekanandan P, Abraham P, Sridharan G, Chandy G, Shaji RV, Daniel D, et al. High frequency of the
precore mutation in patients and blood donors with hepatitis B virus infection from the Indian
subcontinent. Mol Diagn. 2004; 8(1): 51-6. doi: 10.1007/BF03260047. PMID: 15230642.
Funk ML, Rosenberg DM, Lok AS. World‐wide epidemiology of HBeAg‐negative chronic hepatitis B
and associated precore and core promoter variants. J Viral Hepatitis. 2002; 9(1): 52-61. doi:
1046/j.1365-2893.2002.00304.x. PMID: 11851903.
Vray M, Debonne JM, Sire JM, Tran N, Chevalier B, Plantier JC, et al. Molecular epidemiology of
hepatitis B virus in Dakar, Senegal. J Med Virol. 2006; 78(3): 329-34. doi: 10.1002/jmv.20544. PMID:
Biswas A, Chandra PK, Datta S, Panigrahi R, Banerjee A, Chakrabarti S, et al. Frequency and distribution
of hepatitis B virus genotypes among eastern Indian voluntary blood donors: Association with precore and
basal core promoter mutations. Hepatol Res. 2009; 39(1): 53-9. doi: 10.1111/j.1872-034X.2008.00403.x.
PMID: 18713275.
Davidson F, Lycett C, Sablon E, Petrik J, Dow BC. Hepatitis B virus genotypes and precore mutations in
Scottish blood donors. Vox Sang. 2005; 88(2): 87-92. doi: 10.1111/j.1423-0410.2005.00597.x. PMID:
Bläckberg J, Kidd‐Ljunggren K. Genotypic differences in the hepatitis B virus core promoter and precore
sequences during seroconversion from HBeAg to anti‐HBe. J Med Virol. 2000; 60(2): 107-12. doi:
1002/(SICI)1096-9071(200002)60:2<107::AID-JMV1>3.0.CO;2-T. PMID: 10596007.
Lindh M, Andersson AS, Gusdal A. Genotypes, nt 1858 variants, and geographic origin of hepatitis B
virus—large-scale analysis using a new genotyping method. J Infect Dis. 1997; 175(6): 1285-93. doi:
1086/516458. PMID: 9180165.
Chan HL, Hussain M, Lok AS. Different hepatitis B virus genotypes are associated with different
mutations in the core promoter and precore regions during hepatitis B e antigen seroconversion.
Hepatology. 1999; 29(3): 976-84. doi: 10.1002/hep.510290352. PMID: 10051506.
Friedt M, Gerner P, Lausch E, Trübel H, Zabel B, Wirth S. Mutations in the basic core promotor and the
precore region of hepatitis B virus and their selection in children with fulminant and chronic hepatitis B.
Hepatology. 1999; 29(4): 1252-8. doi: 10.1002/hep.510290418. PMID: 10094972.
Laskus T, Rakela J, Persing DH. The stem-loop structure of the cis-encapsidation signal is highly
conserved in naturally occurring hepatitis B virus variants. Virology. 1994; 200(2): 809-12. doi:
1006/viro.1994.1247. PMID: 8178463.
Kramvis A, Kew MC. Relationship of genotypes of hepatitis B virus to mutations, disease progression and
response to antiviral therapy. J Viral Hepatitis. 2005; 12(5): 456-64. doi: 10.1111/j.1365- 2893.2005.00624.x. PMID: 16108759.
Arankalle VA, Murhekar KM, Gandhe SS, Murhekar MV, Ramdasi AY, Padbidri VS, et al. Hepatitis B
virus: predominance of genotype D in primitive tribes of the Andaman and Nicobar islands, India (1989–
. J Gen Virol. 2003; 84(Pt 7): 1915-20. doi: 10.1099/vir.0.18943-0. PMID: 12810887.
Kim H, Jee Y, Mun HS, Song BC, Park JH, Hyun JW, et al. Comparison of full genome sequences between
two hepatitis B virus strains with or without preC mutation (A1896) from a single Korean hepatocellular
carcinoma patient. J Microbiol Biotechnol. 2007; 17(4): 701-4. PMID: 18051288.
Amini‐Bavil‐Olyaee S, Sarrami‐Forooshani R, Mahboudi F, Sabahi F, Adeli A, Noorinayer B, et al.
Genotype characterization and phylogenetic analysis of hepatitis B virus isolates from Iranian patients. J
Med Virol. 2005; 75(2): 227-34. doi: 10.1002/jmv.20261. PMID: 15602742.
Taghavi SA, Tabibi M, Eshraghian A, Keyvani H, Eshraghian H. Prevalence and clinical significance of
hepatitis B Basal core promoter and precore gene mutations in southern Iranian patients. Hepat Mon. 2010;
(4): 294-7. PMID: 22312396, PMCID: PMC3271323.
Halfon P, Bourliere M, Pol S, Benhamou Y, Ouzan D, Rotily M, et al. Multicentre study of hepatitis B
virus genotypes in France: correlation with liver fibrosis and hepatitis B e antigen status. J Viral Hepat.
; 13(5): 329-35. doi: 10.1111/j.1365-2893.2005.00692.x. PMID: 16637864.
Wintermeyer P, Gerner P, Gehring S, Karimi A, Wirth S. Prevalence of hepatitis B virus precore stop
codon mutations in chronically infected children. World J Gastroenterol. 2006; 12(14): 2235-8. doi:
3748/wjg.v12.i14.2235. PMID: 16610027, PMCID: PMC4087652.
Chang MH, Hsu HY, Ni YH, Tsai KS, Lee PI, Chen PJ, et al. Precore stop codon mutant in chronic
hepatitis B virus infection in children: its relation to hepatitis B e seroconversion and maternal hepatitis B
surface antigen. J Hepatol. 1998; 28(6): 915-22. doi: 10.1016/S0168-8278(98)80337-1. PMID: 9672164.
Gandhe SS, Chadha MS, Arankalle VA. Hepatitis B virus genotypes and serotypes in western India: lack of
clinical significance. J Med Virol. 2003; 69(3): 324-30. doi: 10.1002/jmv.10292. PMID: 12526041.
Abbas Z, Muzaffar R, Siddiqui A, Naqvi SA, Rizvi SA. Genetic variability in the precore and core
promoter regions of hepatitis B virus strains in Karachi. BMC Gastroenterol. 2006; 6: 20. doi:
1186/1471-230X-6-20. PMID: 16863587, PMCID: PMC1544342.
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