Indian Journal of Medical Microbiology IAMM  | About us |  Subscription |  e-Alerts  | Feedback |  Login   
  Print this page Email this page   Small font sizeDefault font sizeIncrease font size
 Home | Ahead of Print | Current Issue | Archives | Search | Instructions  
Users Online: 293 Official Publication of Indian Association of Medical Microbiologists 
  Search
 
 ~ Next article
 ~ Previous article 
 ~ Table of Contents
  
 ~  Similar in PUBMED
 ~  Search Pubmed for
 ~  Search in Google Scholar for
 ~Related articles
 ~  Article in PDF (345 KB)
 ~  Citation Manager
 ~  Access Statistics
 ~  Reader Comments
 ~  Email Alert *
 ~  Add to My List *
* Registration required (free)  

 
 ~  Abstract
 ~  Introduction
 ~  Materials and Me...
 ~  Results
 ~  Discussion
 ~  Conclusion
 ~  Acknowledgments
 ~  References
 ~  Article Figures
 ~  Article Tables

 Article Access Statistics
    Viewed4988    
    Printed244    
    Emailed3    
    PDF Downloaded270    
    Comments [Add]    
    Cited by others 15    

Recommend this journal

 


 
BRIEF COMMUNICATION
Year : 2010  |  Volume : 28  |  Issue : 1  |  Page : 54-56
 

Determination of hepatitis C virus genotypes among blood donors in Ahvaz, Iran


1 Department of Virology, Faculty of Medicine and Tropical and Infectious Disease Research Centre, Ahvaz Jondishapour University of Medical Sciences, Golestan Blved, Ahvaz, Iran
2 Ahvaz Blood Transfusion Centre, Ahvaz, Iran

Date of Submission22-Apr-2009
Date of Acceptance28-Jul-2009
Date of Web Publication6-Jan-2010

Correspondence Address:
F Farshadpour
Department of Virology, Faculty of Medicine and Tropical and Infectious Disease Research Centre, Ahvaz Jondishapour University of Medical Sciences, Golestan Blved, Ahvaz
Iran
Login to access the Email id

Source of Support: Ahvaz Jondishapur University of Medical Science, Conflict of Interest: None


DOI: 10.4103/0255-0857.58731

Rights and Permissions

 ~ Abstract 

This study aims to determine the genotypes of hepatitis C virus (HCV) among blood donors at Ahvaz Blood Transfusion Centre. Blood samples were taken from 2376 blood donors - 1795 (75.54%) male and 581(24.45%) female - who referred to Ahvaz Blood Transfusion Centre during 2007-2008. Detection of anti-HCV antibody for all the donors was carried out by ELISA and the confirmatory RIBA tests. HCV RT-PCR followed by RFLP test was carried out for anti-HCV positive samples. Out of 2376 blood donors, only 55 (2.3%) male donors showed to be positive for HCV antibody by ELISA and RIBA tests out of which 45(1.8%) donors were positive for RT-PCR test. Female donors were negative for HCV antibody. The result of HCV genotyping by RFLP test showed 24 (53.3%) for 1a, 17 (37.7%) for 3a (a) and 4 (8.8%) for 3a (b) genotypes respectively. In conclusion, high prevalence of 53.3% HCV 1a genotype was observed among blood donors in Ahvaz city.


Keywords: Blood donors, genotyping, Hepatitis C virus, Restriction Fragment Length Polymorphism, RT-PCR


How to cite this article:
Farshadpour F, Makvandi M, Samarbafzadeh A R, Jalalifar M A. Determination of hepatitis C virus genotypes among blood donors in Ahvaz, Iran. Indian J Med Microbiol 2010;28:54-6

How to cite this URL:
Farshadpour F, Makvandi M, Samarbafzadeh A R, Jalalifar M A. Determination of hepatitis C virus genotypes among blood donors in Ahvaz, Iran. Indian J Med Microbiol [serial online] 2010 [cited 2019 Oct 16];28:54-6. Available from: http://www.ijmm.org/text.asp?2010/28/1/54/58731



 ~ Introduction Top


Hepatitis C virus (HCV) is a single stranded RNA virus which belongs to the Flaviviridae family. [1] HCV is a causative agent for chronic, acute and fulminant hepatitis. [2],[3] The association of HCV among patients with cirrhosis of liver and hepatocellular carcinoma has been reported. [4] HCV infection is a global health problem and it is estimated that 200 million people of the world population are infected with HCV. [5]

The study of genetic variability of HCV strains has led the consensus classification into six major genotypes [Table 1]. [3] Some studies suggest that the clinical features of liver disease depend on HCV genotypes. [6] It is also noteworthy that the success of Interferon and Ribavirin treatment seems to be related to genotypes. [7] Further, HCV genotyping is a useful tool to determine its molecular epidemiology. [2] Ahvaz city is the centre of Khozestan province with a population of about 1.8 million located in the South-West of Iran. There has been no data available so far on HCV genotyping among blood donors in Ahvaz city. This study was conducted to determine HCV genotypes among blood donors in Ahvaz city.


 ~ Materials and Methods Top


This study, (with research project number U-86069,) was approved by the ethical committee of Ahvaz Jondishapur University of Medical Science. All blood donors gave consent to take part in the experiments voluntarily.

The sera were collected from 2376 blood donors referred to the Blood Transfusion Centre from August 2007 to February 2008. The average age was varying from 19 to 50 while the mean age was 34.5.

Serological data

The sera were collected from these 2376 blood donors. All the sera were initially tested for anti-HCV antibody by Enzyme Linked Immuno sorbent Assay (ELISA) test (Ortho; HCV 3.0 ELISA Test System; Ortho-Clinical Diagnostics, Raritan, NJ, USA). For positive sera by ELISA, the confirmatory Recombinant Immunoblot Assay (RIBA) test (Deciscan; HCV plus; Sanofi Diagnostics Pasteur, SA Marnes la Coquuette, France) was carried out. [2]

The positive HCV Ab sera were aliquoted and stored immediately at minus 70C before use.

Extraction of hepatitis C virus RNA

We mixed 250 micro litres of serum with 750 ΅l of Trizol LS (Trizol; LS Reagent: Invitrogen, Life Technologies, Carlsbad, CA, USA) for the HCV RNA extraction followed by precipitating by ethanol and then drying.

cDNA synthesis

HCV-RNA was immediately transcribed into cDNA using random primers (Sensicript RT kit; QIAGEN, Poison Information Centre Mainz, Germany). [8]

Hepatitis C virus-RNA detection

cDNA was targeted by a nested-PCR directed at the 5' untranslated region using two sets of primers (ROCHE; Roche Diagnostics GmbH, Roche Applied Science, Mannheim, Germany), the first set 5`-AGC GTC TAG CCA TGG CGT -3`, called forward external primer, and 5`- GCA CGG TCT ACG AGA CCT-3`, named reverse external primer. The second set 5`-GTG GTC TGC GGA ACC GG-3`, called forward inner primer, and 5`- GGG CAC TCG CAA GCA CCC-3`, named reverse inner primer of 5'UTR region of HCV genome. The first round was carried out for 30 cycles which consisted of initial denaturation at 94C for 5 min, denaturation at 94C for 35sec, annealing at 58C for 40sec, extension at 72C for 45sec and, the final extension at 72C for 5 min. The second round was followed for 25 cycles which consisted of initial denaturation at 94C for five minutes, denaturation at 94C for 35 seconds, annealing at 64C for 40 seconds, extension at 72C for 45 seconds, and the final extension at 72C for five minutes. The 174-bp second PCR product was submitted to electrophoresis by using a 1.5% agarose gel in 0.5X TBE buffer, and was visualized by ethidium bromide staining under ultraviolet light. [8],[9]

Genotyping

The amplicons of each sample were digested by the following three sets of restriction enzymes at 37C for three hours:

  1. Scrf I/Hinf I
  2. MvaI/HinfI
  3. MvnI.
Then the genotypes 1a, 1b, 2a, 2b, 3a, 3b, [4],[5] and 6 were distinguishable. [2],[8],[9]


 ~ Results Top


Out of 2376 donors, 1795 (75.54%) were males and 581 (24.45%) were females.

Of the 2376 blood donors, 55 (2.3%) male donors showed to be positive for HCV antibody while only 45 (1.8%) samples were positive by RT-PCR test. [Figure 1] shows the 174 bp PCR product. The result of HCV genotyping by RFLP test revealed 24 (53.3%) for 1a, 17 (37.7%) for 3a (a), and 4(8.8%) for 3a (b) genotypes, respectively. [Figure 2] shows the pattern of RFLP results.


 ~ Discussion Top


Epidemiological studies in different regions of the world show the wide variation in HCV prevalence patterns [5] though it is clearly revealed that the incidence of HCV is higher among less developed nations. [5] In Iran, HCV prevalence in general population is less than one per cent. [4] In our study, the prevalence of hepatitis C infection was about 2.3% by RIBA and 1.8% by PCR tests, respectively. Regarding the HCV genotypes, 24(53.3%) for 1a, 17 (37.7%) for 3a (a) and 4(8.8%) for 3a (b) genotypes were found among the blood donors. Our results are nearly similar to those studies which have already been conducted by different investigators in Tehran- the capital of Iran. [10],[11]

Regarding HCV genotypes, Samimi Rad et al., found 1a (47%), 3a (36%), 1b (8%), and four (seven per cent) genotypes among Iranian patients, and revealed that genotype 1a is frequent in the South of Iran (70%), while 3a is more prevalent in the North-West of Iran (83%). Patients infected by blood products more frequently had genotype 1a (57%), while younger drug users had genotype 3a (54%) more frequently. [10]

Keyvani et al., studied distribution frequency of hepatitis C virus genotypes in Iran. In their study, genotype 1a with 39.7% had the highest frequency. Genotype 3a (27.5%) and 1b (12.1%) were the other frequent genotypes. [11]

The prevalence of hepatitis C among blood donors is less than one per cent in Northern European countries. [5] Higher rates have been reported in South East Asian countries, including India (1.5%), Malaysia (2.3%), and the Philippines (2.3%). [5] The incidence in Japan is 1.2%. [5] Alarming rates of 14.5% are reported in Egypt. [5] HCV genotype 4 is common in countries such as Yemen, Kuwait, Iraq, and Saudi Arabia. [12] However, genotype 1b is prevalent in Turkey, [12] and HCV genotype 3a is prevalent in Pakistan. [13] Genotype information is important when HCV treatment is being considered, since some genotypes respond more favourably to the medications. Genotype also determines the length of therapy e.g., treatment for genotypes 2 and 3 requires only 24 weeks while genotypes 1 and 4 require 48 weeks. [6] In addition to treatment purposes, detection of HCV genotypes in different regions can be used for the purpose of molecular epidemiology. [6]

The results of this study have some similarities with other studies in Pakistan and India, which show that genotype 3 is very prevalent and genotype 2 is very rare. [13] Other studies have shown the absence of genotype 2 in Iran. [13]


 ~ Conclusion Top


High prevalence of 53.3% 1a genotype and low prevalence of 8.8% 3a (b) HCV genotypes were observed among blood donors in Ahvaz city.


 ~ Acknowledgments Top


The authors would like to thank Deputy Research and Affairs of Ahvaz Jondishapur University of Medical Science for financial support to meet all expenses and essential equipment for this study.

 
 ~ References Top

1.Choo QL, Kuo G, Weiner AJ, et al. Isolation of a cDNA clone derived from a blood-borne non-A, non-B viral hepatitis genome. Science 1989;244:359-62.  Back to cited text no. 1      
2.Silva LK, Silva MB, Rodart IF, Lopes GB, Costa FQ, Melo ME, et al. Prevalence of hepatitis C virus (HCV) infection and HCV genotypes of hemodialysis patients in Salvador, Northeastern Brazil. Braz J Med Biol Res 2006;39:595-602.  Back to cited text no. 2      
3.Alavian SM, Gholami B, Masarrat S. Hepatitis C risk factor in Iranian volunteer blood donors: A case-control study. J Gastrol Hepatol 2002;17:1092-7.  Back to cited text no. 3      
4.Alavian SM. Hepatitis C, Chronic renal failure, control is possible. Hepatitis Monthly 2006;6:51-2.  Back to cited text no. 4      
5.Koop CE. Hepatitis C: An epidemic for any one, worldwide prevalence. DHMC 2008. Available from: http://www.epidemic.org/theFacts/theEpidemic/worldprevalence/. [cited in 2008].  Back to cited text no. 5      
6.Lee CM, Hung CH, Lu SN, Changchien CS. Hepatitis C virus genotypes: Clinical relevance and therapeutic implications. Chang Gung Med J 2008;31:16-25.  Back to cited text no. 6  [PUBMED]  [FULLTEXT]  
7.Senecal DL, Morelli J. Hepatitis C virus infection: A current review. JAAPA 2007. Available from: http://www.jaapa.com/issues/j20071001/articles/hepatitis1007.htm. [cited in 2007].   Back to cited text no. 7      
8.Sambrook J., Russell D. Molecular cloning: A laboratory manual. 3 rd ed. CSHL Press; 2001.  Back to cited text no. 8      
9.Pohjanpelto P. et al. Hepatitis C genotypes in Finland determined by RFLP. Clin Diag Virol 1996;7:7-16.  Back to cited text no. 9      
10.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;74:246-52.  Back to cited text no. 10  [PUBMED]  [FULLTEXT]  
11.Keyvani H, Alizadeh AH, Alavian SM, Ranjbar M, Hatami S. Distribution frequency of hepatitis C virus genotypes in 2231 patients in Iran. Hepatol Res 2007;37:101-3.  Back to cited text no. 11  [PUBMED]  [FULLTEXT]  
12.Simmonds P, Holmes EC, Cha TA, et al. Classification of hepatitis C virus into six major genotypes and a series of subtypes by phylogenetic analysis of the NS-5 region. J Gen Virol 1993;74:2391-9.  Back to cited text no. 12      
13.Chowdhury A, Santra A, Chaudhuri S, Dhali GK, Chaudhuri S, Maity SG, et al. Hepatitis C virus infection in the general population: A community-based study in West Bengal, India. Hepatology 2003;37:802-9.  Back to cited text no. 13      


    Figures

  [Figure 1], [Figure 2]
 
 
    Tables

  [Table 1]

This article has been cited by
1 Prevalence of Hepatitis C Virus Genotypes in Iranian Patients: a Systematic Review and Meta-Analysis
Mazaher Khodabandehloo,Daem Roshani
Hepatitis Monthly. 2014; 14(11)
[Pubmed] | [DOI]
2 Distribution of Hepatitis C Virus Genotypes in Iranian Chronic Infected Patients
Fatemeh Jahanbakhsh Sefidi,Hossein Keyvani,Seyed Hamidreza Monavari,Seyed Moayed Alavian,Shahin Fakhim,Farah Bokharaei-Salim
Hepatitis Monthly. 2013; 13(1)
[Pubmed] | [DOI]
3 Hepatitis B and C Infections and Different Genotypes of HCV Among Sickle Cell Anemia Patients in Ahvaz, South-Western Iran
Mehri Ghafourian-Boroujerdnia,Mohammad Ali Assarehzadegan,Khodamorad Zandian
Jundishapur Journal of Microbiology. 2013; 6(5)
[Pubmed] | [DOI]
4 Distribution of hepatitis C virus genotypes in Iranian chronic infected patients
Sefidi, F.J. and Keyvani, H. and Monavari, S.H. and Alavian, S.M. and Fakhim, S. and Bokharaei-Salim, F.
Hepatitis Monthly. 2013; 13(1)
[Pubmed]
5 Prevalence of hepatitis C virus genotypes in the Northern of Iran (Mazandaran) from 2009 to 2011
Haghshenas, M. and Babamahmoodi, F. and Rafiei, A. and Vahedi, V. and Alizadeh-Navaei, R.
HealthMED. 2012; 6(9): 3046-3050
[Pubmed]
6 Prevalence of hepatitis B and C infections and HCV genotypes among haemophilia patients in Ahvaz, Southwest Iran
Assarehzadegan, M.A. and Boroujerdnia, M.G. and Zandian, K.
Iranian Red Crescent Medical Journal. 2012; 14(8): 3
[Pubmed]
7 Hepatitis C virus infection in the Middle East and North Africa MENA region: injecting drug users (IDUs) is an under-investigated population
S. Ramia,N. M. Melhem,K. Kreidieh
Infection. 2012; 40(1): 1
[Pubmed] | [DOI]
8 Hepatitis C virus infection in the Middle East and North Africa "mENA" region: Injecting drug users (IDUs) is an under-investigated population
Ramia, S., Melhem, N.M., Kreidieh, K.
Infection. 2012; 40(1): 1-10
[Pubmed]
9 Determination of hepatitis C virus genotypes among HCV positive patients in Shahrekord, Iran
Tajbakhsh, E., Dosti, A., Tajbakhsh, S., Momeni, M., Tajbakhsh, F.
# African Journal of Microbiology Research. 2011; 5(32): 5910-5915
[Pubmed]
10 The correlation between HCV genotypes and liver damage in HCV patients
Haghshenas, M., Babamahmoodi, F., Rafiee, A., Vahedi, V.
Journal of Mazandaran University of Medical Sciences. 2011; 21(85): 75-83
[Pubmed]
11 Seroprevalence of hepatitis C virus in diagnostic patients and blood donors in and around davangere
Raghu Kumar, K.G., Eshwar Singh, R., Chandrappa, N.R., Hanumanthappa, A.R., Vijayanath, V., Anitha, M.R.
Journal of Pure and Applied Microbiology. 2011; 5(1): 275-279
[Pubmed]
12 Infection of hepatitis C virus genotypes in hepatocellular carcinoma patients from rural areas of Faisalabad region, Pakistan
Nawaz, M., Siddique, S., Manzoor, I., Nadeem, S., Salah-ud-Din, Shabbir, H., Naveed, R., Rajoka, M.I.
African Journal of Biotechnology. 2011; 10(42): 8471-8475
[Pubmed]
13 Detection of hepatitis C virus and antibodies in postmortem blood and bloodstains
Takasaka, T., Itoh, Y., Kaneko, H., Ikegaya, H.
Journal of Clinical Microbiology. 2011; 49(3): 1122-1123
[Pubmed]
14 Molecular epidemiology of hepatitis C virus genotypes in Bushehr province, Iran
Vahdat, K., Keyvani, H., Tabib, S.M., Rostamabadi, S., Valizadeh, S.M., Cheraghi, S., Shamsian, S., Zandi, K.
European Review for Medical and Pharmacological Sciences. 2010; 14(10): 861-864
[Pubmed]
15 Viral hepatitis in patients hospitalized in two teaching hospitals, Tehran, Iran
Talebi-Taher, M., Mohit, M., Avanessians, T.B.
Iranian Journal of Clinical Infectious Diseases. 2010; 5(1): 25-29
[Pubmed]



 

Top
Print this article  Email this article
Previous article Next article

    

2004 - Indian Journal of Medical Microbiology
Published by Wolters Kluwer - Medknow

Online since April 2001, new site since 1st August '04