شناسایی جهش های ژن NS5B ژنوتیپ 1a ویروس هپاتیت C در استان گیلان
محورهای موضوعی : ویروس شناسی
لیلا اسدپور
1
(استادیار، گروه زیست شناسی، واحد رشت، دانشگاه آزاد اسلامی، رشت، ایران.)
حمدالله رحیمی دوگاهه
2
(کارشناس ارشد، گروه زیست شناسی، واحد تنکابن، دانشگاه آزاد اسلامی، تنکابن، ایران.)
مهناز هوشمند راد
3
(کارشناس ارشد، گروه زیست شناسی، واحد تنکابن، دانشگاه آزاد اسلامی، تنکابن، ایران.)
کلید واژه: مقاومت دارویی, هپاتیت C, جهش NS5B,
چکیده مقاله :
سابقه و هدف: ویروس هپاتیت C عامل اصلی هپاتیت مزمن کبدی است که سالیانه باعث مرگ هزاران نفر در دنیا می گردد. پروتئین NS5B، RNA پلی مراز وابسته به RNA است که توسط ژن NS5B کد می شود و در همانندسازی ویروس نقش دارد. از جمله داروهای موثر در درمان این عفونت های ناشی از این ویروس مهارکننده های پروتئین NS5B می باشند. ظهور سویه های مقاوم به این داروها یک مانع بزرگ در موفقیت درمان می باشد. هدف از این مطالعه بررسی جهش های احتمالی در ناحیه NS5B ژنوتیپ 1a ویروس هپاتیت C در استان گیلان است.مواد و روش ها: RNA ژنومی ویروس از پلاسمای 225 بیمار آنتیHCV مثبت استخراج و شناسایی مولکولی و تعیین سروتیپ آن به روشRT-PCR و تعیین توالی محصول انجام شد. پس از آن ژن NS5B در 10 سویه دارای ژنوتیپ 1a تکثیر و به منظور شناسایی جهش ها توالی یابی گردید.یافته ها: به ترتیب ژنوتیپ های 3a (53.3 درصد) و 1a (36.9 درصد) فراوان ترین ژنوتیپ های شناسایی شده در گیلان بودند. بر اساس نتایج توالی یابی، از میان 10 سویه دارای ژنوتیپ 1a مورد بررسی در 5 سویه ، 7 نوع جهش بدمعنی در کدون های Q309، A327، S254، K304، N307، R250 و A334 شناسایی شد.نتیجه گیری: ژنوتیپ 1a از ژنوتیپ های شایع ویروس هپاتیت C در گیلان می باشد. شناسایی جهش ها یا پلی مورفیسم مرتبط با مقاومت در ویروس هپاتیت C می تواند در بهینه سازی درمان و تعیین کارایی داروها در درمان هپاتیت C مفید باشد.
Background & Objectives: Hepatitis C virus is the main cause of chronic hepatitis, resulting in a thousand deaths every year. NS5B protein is an RNA-dependent RNA polymerase that is encoded by the NS5B gene and involved in virus replication. One of the most effective drugs in the treatment of infections caused by this virus are NS5B protein inhibitors. The emergence of the strains resistant to these drugs is a major obstacle to the success of treatment. The aim of this study was to investigate possible mutations in the NS5B region of hepatitis C virus genotype 1a in Guilan province. Materials & Methods: HCV genomic RNA was extracted from the plasma samples of 225 anti-HCV positive patients and its molecular identification and genotyping was carried out by RT-PCR and product Sequencing. Then, the NS5B gene was amplified in 10 strains with genotype 1a and subsequently sequenced to determine mutations in this region. Results: Genotypes 3a (53.3%) and 1a (36.9%) were the most abundant genotypes identified in Guilan. According to sequencing results, out of 10 investigated genotype 1a strains, 5 strains showed 7 types of missense mutations in codons Q309, A327, S254, K304, N307, R250, and A334. Conclusions: Genotype 1a is one of the common genotypes of hepatitis C virus in Guilan. Identifying mutations or polymorphisms associated with resistance in hepatitis C virus can be useful in optimizing the treatment and determining the efficacy of drugs in treating hepatitis C virus.
Klenerman P, Chinnakannan S, Barnes E. The generation of a simian adenoviral vectored HCV
vaccine encoding genetically conserved gene segments to target multiple HCV genotypes.
Vaccine. 2018; 36(2): 313-321.
2. Chantratita W, Song KS, Gun Ho C, Pongthanapisith V, Thongbaiphet N, Wongtabtim G,
Pasomsub E, Angkanavin K, Nimse SB, Sonawane MD, Warkad SD. 6 HCV genotyping 9G test
and its comparison with VERSANT HCV genotype 2.0 assay (LiPA) for the hepatitis C virus
genotyping. J Virol Methods. 2017; 239: 1-8.
3. Ali N, Allam H, May R, Sureban SM, Bronze MS, Bader T, Umar S, Anant S, Houchen CW.
Hepatitis C virus-induced cancer stem cell-like signatures in cell culture and murine tumor
xenografts. J Virol. 2011; 85(23): 12292-12303.
4. Messina JP, Humphreys I, Flaxman A, Brown A, Cooke GS, Pybus OG, Barnes E. Global
distribution and prevalence of hepatitis C virus genotypes. Hepatol. 2015; 61(1): 77-87.
5. Lauer GM, Walker BD. Hepatitis C virus infection. New Engl J Med. 2001; 345(1): 41-52.
6. Bennett JE, Dolin R, Blaser MJ. Principles and practice of infectious diseases. Elsevier Health
Sci; 2014; 1: 1336-1356.
7. Ranjith-Kumar CT, Kao CC. Biochemical activities of the HCV NS5B RNA-dependent RNA
polymerase. Hepatitis C viruses: Genomes and molecular biology. Norfolk (UK): Horizon
Bioscience. 2006: 293-310.
8. Pockros PJ. New direct-acting antivirals in the development for hepatitis C virus infection.
Therap Adv Gastroenterol. 2010; 3(3): 191-202.
9. Motawi TM, Rizk SM, Shaker OG, Mokhtar OZ. MicroRNAs as predictor markers for response
to interferon treatment of chronic hepatitis C genotype-4 in Egyptian patients. PloS one. 2015; 10
(3): 1-12.
10. Castilho MC, Martins AN, Horbach IS, Perez RD, Figueiredo FA, Pinto PD, Nabuco LC, Lima
DB, Tanuri A, Porto LC, Júnior F. Association of hepatitis C virus NS5B variants with resistance
to new antiviral drugs among untreated patients. Mem Inst Oswaldo Cruz. 2011; 106(8):
968-975.
11. Huang T, Wang J, Cai YD, Yu H, Chou KC. Hepatitis C virus network based classification of
hepatocellular cirrhosis and carcinoma. PLoS One. 2012; 7(4): e34460.
12. Hosseini Moghaddam SM, Keyvani H, Kasiri H, Kazemeyni SM, Basiri A, Aghel N, Alavian
SM. Distribution of hepatitis C virus genotypes among hemodialysis patients in Tehran: a
multicenter study. J Med Virol. 2006; 78(5): 569-573.
13. Verma V, Chakravarti A, Kar P. Genotypic characterization of hepatitis C virus and its
significance in patients with chronic liver disease from Northern India. Diagn Microbiol Infect
Dis. 2008; 61(4): 408-414.
14. Haghshenas MR, Babamahmoodi F, Rafiei AR, Vahedi V. The correlation between HCV
genotypes and liver damage in HCV patients. J Mazandaran Uni Med Sci. 2011; 21(85): 76-83.
15. Samimi Rad K, Nategh R, Malekzadeh R, Norder H, Magnius L. Molecular epidemiology of
hepatitis C virus in Iran as reflected by phylogenetic analysis of the NS5B region. J Med Virol.
2004; 74(2): 246-252.
16. Makhloogh A, Aezinia N, Haghshenas MR, Tirgar-fakheri H, Maleki I, Taghvaei T.
Comparison of Hepatitis C Virus Genotypes in Hemodialysis and Nonuremic Patients.
Armaghane Danesh. 2010; 15(3): 283-292. [In Persian]
17. Hejazi MS, Ghotaslou R, Farshdoosty Hagh M, Mohammadzadeh Sadigh Y. Genotyping of
hepatitis C virus in northwest of Iran. Biotechnol. 2007; 6(3): 302-308.
18. Jaspe RC, Sulbarán YF, Sulbarán MZ, Loureiro CL, Rangel HR, Pujol FH. Prevalence of
amino acid mutations in hepatitis C virus core and NS5B regions among Venezuelan viral
isolates and comparison with worldwide isolates. Virol J. 2012; 9(1): 214.
19. Kim DW, Lee SA, Kim H, Won YS, Kim BJ. Naturally occurring mutations in the
nonstructural region 5B of hepatitis C virus (HCV) from treatment-naïve Korean patients
chronically infected with HCV genotype 1b. PLOS One. 2014; 9(1): e87773.
20. Paolucci S, Fiorina L, Mariani B, Gulminetti R, Novati S, Barbarini G, Bruno R, Baldanti F.
Naturally occurring resistance mutations to inhibitors of HCV NS5A region and NS5B
polymerase in DAA treatment-naive patients. Virol J. 2013; 10(1): 355.
_||_
Klenerman P, Chinnakannan S, Barnes E. The generation of a simian adenoviral vectored HCV
vaccine encoding genetically conserved gene segments to target multiple HCV genotypes.
Vaccine. 2018; 36(2): 313-321.
2. Chantratita W, Song KS, Gun Ho C, Pongthanapisith V, Thongbaiphet N, Wongtabtim G,
Pasomsub E, Angkanavin K, Nimse SB, Sonawane MD, Warkad SD. 6 HCV genotyping 9G test
and its comparison with VERSANT HCV genotype 2.0 assay (LiPA) for the hepatitis C virus
genotyping. J Virol Methods. 2017; 239: 1-8.
3. Ali N, Allam H, May R, Sureban SM, Bronze MS, Bader T, Umar S, Anant S, Houchen CW.
Hepatitis C virus-induced cancer stem cell-like signatures in cell culture and murine tumor
xenografts. J Virol. 2011; 85(23): 12292-12303.
4. Messina JP, Humphreys I, Flaxman A, Brown A, Cooke GS, Pybus OG, Barnes E. Global
distribution and prevalence of hepatitis C virus genotypes. Hepatol. 2015; 61(1): 77-87.
5. Lauer GM, Walker BD. Hepatitis C virus infection. New Engl J Med. 2001; 345(1): 41-52.
6. Bennett JE, Dolin R, Blaser MJ. Principles and practice of infectious diseases. Elsevier Health
Sci; 2014; 1: 1336-1356.
7. Ranjith-Kumar CT, Kao CC. Biochemical activities of the HCV NS5B RNA-dependent RNA
polymerase. Hepatitis C viruses: Genomes and molecular biology. Norfolk (UK): Horizon
Bioscience. 2006: 293-310.
8. Pockros PJ. New direct-acting antivirals in the development for hepatitis C virus infection.
Therap Adv Gastroenterol. 2010; 3(3): 191-202.
9. Motawi TM, Rizk SM, Shaker OG, Mokhtar OZ. MicroRNAs as predictor markers for response
to interferon treatment of chronic hepatitis C genotype-4 in Egyptian patients. PloS one. 2015; 10
(3): 1-12.
10. Castilho MC, Martins AN, Horbach IS, Perez RD, Figueiredo FA, Pinto PD, Nabuco LC, Lima
DB, Tanuri A, Porto LC, Júnior F. Association of hepatitis C virus NS5B variants with resistance
to new antiviral drugs among untreated patients. Mem Inst Oswaldo Cruz. 2011; 106(8):
968-975.
11. Huang T, Wang J, Cai YD, Yu H, Chou KC. Hepatitis C virus network based classification of
hepatocellular cirrhosis and carcinoma. PLoS One. 2012; 7(4): e34460.
12. Hosseini Moghaddam SM, Keyvani H, Kasiri H, Kazemeyni SM, Basiri A, Aghel N, Alavian
SM. Distribution of hepatitis C virus genotypes among hemodialysis patients in Tehran: a
multicenter study. J Med Virol. 2006; 78(5): 569-573.
13. Verma V, Chakravarti A, Kar P. Genotypic characterization of hepatitis C virus and its
significance in patients with chronic liver disease from Northern India. Diagn Microbiol Infect
Dis. 2008; 61(4): 408-414.
14. Haghshenas MR, Babamahmoodi F, Rafiei AR, Vahedi V. The correlation between HCV
genotypes and liver damage in HCV patients. J Mazandaran Uni Med Sci. 2011; 21(85): 76-83.
15. Samimi Rad K, Nategh R, Malekzadeh R, Norder H, Magnius L. Molecular epidemiology of
hepatitis C virus in Iran as reflected by phylogenetic analysis of the NS5B region. J Med Virol.
2004; 74(2): 246-252.
16. Makhloogh A, Aezinia N, Haghshenas MR, Tirgar-fakheri H, Maleki I, Taghvaei T.
Comparison of Hepatitis C Virus Genotypes in Hemodialysis and Nonuremic Patients.
Armaghane Danesh. 2010; 15(3): 283-292. [In Persian]
17. Hejazi MS, Ghotaslou R, Farshdoosty Hagh M, Mohammadzadeh Sadigh Y. Genotyping of
hepatitis C virus in northwest of Iran. Biotechnol. 2007; 6(3): 302-308.
18. Jaspe RC, Sulbarán YF, Sulbarán MZ, Loureiro CL, Rangel HR, Pujol FH. Prevalence of
amino acid mutations in hepatitis C virus core and NS5B regions among Venezuelan viral
isolates and comparison with worldwide isolates. Virol J. 2012; 9(1): 214.
19. Kim DW, Lee SA, Kim H, Won YS, Kim BJ. Naturally occurring mutations in the
nonstructural region 5B of hepatitis C virus (HCV) from treatment-naïve Korean patients
chronically infected with HCV genotype 1b. PLOS One. 2014; 9(1): e87773.
20. Paolucci S, Fiorina L, Mariani B, Gulminetti R, Novati S, Barbarini G, Bruno R, Baldanti F.
Naturally occurring resistance mutations to inhibitors of HCV NS5A region and NS5B
polymerase in DAA treatment-naive patients. Virol J. 2013; 10(1): 355.