Determination of an individual’s hepatitis C virus (HCV) genotypes prior to antiviral therapy has become increasingly important for the clinical management and prognosis of HCV infection. Therefore, this study was conducted to investigate the prevalence of HCV genotypes in HCV infected patients of district Bannu in Khyber Pakhtunkhwa region of Pakistan. Serum samples of 117 seropositive patients were screened for HCV-RNA by using reverse transcriptase-nested polymerase chain reaction (RT-nested PCR) and then PCR positive samples were subjected to HCV genotyping. Out of 117 seropositive samples, 110 samples were found positive by PCR analysis. Genotype 3a was the most prevalent one detected in 38% of patients, followed by genotype 3b in 21% of patients, and then genotype 2a in 12% of patients. However 21% of HCV-PCR positive samples could not be genotyped by method used in this study. Genotype 3a was the most prevalent genotype in patients of all age groups and its prevalence was found high among patients with increasing age (>34 years). Moreover, genotypes 3a and 3b were found to be the most prevalent genotypes in patients with history of shaving by barbers, receiving multiple injections, and dental procedures. In conclusion there is need of further investigation of genotypes of HCV by using more sensitive assays and considering large sample size in district Bannu. 1. Introduction HCV infection is among life threatening public health problems worldwide, with over 170–200 million infected people [1] including about 17 million from Pakistan [2]. HCV is considered the leading cause of liver cirrhosis and hepatocellular carcinoma. It has been estimated to cause approximately 27% of cirrhosis and 25% of hepatocellular carcinoma cases worldwide [3]. Each year about 350,000 people die due to HCV [4]. HCV is a small enveloped, positive sense single stranded RNA virus and has been classified as a separate genus hepacivirus in the Flaviviridae family [5]. The HCV genome is approximately 9.6？kb, encoding a polyprotein of about 3010 amino acids and is flanked by short untranslated regions (UTRs) regions at the 5′ and 3′ terminus [6]. This polyprotein is posttranslationally processed by viral and cellular proteins to generate the structural proteins (C, E1, E2, and p7) and nonstructural proteins (NS2, NS3, NS4A, NS4B, NS5A, and NS5B), [7]. HCV shows high degree of genetic heterogeneity; consequently six major genotypes and multiple subtypes of HCV have been identified so far in world [8]. Distribution of HCV genotypes and subtypes in different regions of the world
References
[1]
S. Butt, M. Idrees, H. Akbar et al., “The changing epidemiology pattern and frequency distribution of hepatitis C virus in Pakistan,” Infection, Genetics and Evolution, vol. 10, no. 5, pp. 595–600, 2010.
[2]
M. Idrees, S. Rafique, I. Rehman et al., “Hepatitis C virus genotype 3a infection and hepatocellular carcinoma: Pakistan experience,” World Journal of Gastroenterology, vol. 15, no. 40, pp. 5080–5085, 2009.
[3]
M. J. Alter, “Epidemiology of hepatitis C virus infection,” World Journal of Gastroenterology, vol. 13, no. 17, pp. 2436–2441, 2007.
[4]
A. Hatzakis, S. Wait, J. Bruix et al., “The state of hepatitis B and C in Europe: report from the hepatitis B and C summit conference,” Journal of Viral Hepatitis, vol. 18, no. 1, pp. 1–16, 2011.
[5]
C. H. Hagedorn, E. H. van Beers, and C. de Staercke, “Hepatitis C virus RNA-dependent RNA polymerase (NS5B polymerase),” Current Topics in Microbiology and Immunology, vol. 242, p. 327, 2000.
[6]
N. Kato, “Molecular virology of hepatitis C virus,” Acta Medica Okayama, vol. 55, no. 3, pp. 133–159, 2001.
[7]
M. Liew, M. Erali, S. Page, D. Hillyard, and C. Wittwer, “Hepatitis C genotyping by denaturing high-performance liquid chromatography,” Journal of Clinical Microbiology, vol. 42, no. 1, pp. 158–163, 2004.
[8]
M. Idrees, A. Lal, M. Naseem, and M. Khalid, “High prevalence of hepatitis C virus infection in the largest province of Pakistan,” Journal of Digestive Diseases, vol. 9, no. 2, pp. 95–103, 2008.
[9]
A. R. N. Zekri, A. A. Bahnassy, H. M. A. El-Din, and H. M. Salama, “Consensus siRNA for inhibition of HCV genotype-4 replication,” Virology Journal, vol. 6, article 13, 2009.
[10]
R. W. Chamberlain, N. Adams, A. A. Saeed, P. Simmonds, and R. M. Elliott, “Complete nucleotide sequence of a type 4 hepatitis C virus variant, the predominant genotype in the Middle East,” Journal of General Virology, vol. 78, no. 6, pp. 1341–1347, 1997.
[11]
A. S. Abdulkarim, N. N. Zein, J. J. Germer et al., “Hepatitis C virus genotypes and hepatitis G virus in hemodialysis patients from Syria: identification of two novel hepatitis C virus subtypes,” The American Journal of Tropical Medicine and Hygiene, vol. 59, no. 4, pp. 571–576, 1998.
[12]
N. N. Zein, “Clinical significance of hepatitis C virus genotypes,” Clinical Microbiology Reviews, vol. 13, no. 2, pp. 223–235, 2000.
[13]
D. Amarapurkar, M. Dhorda, A. Kirpalani, A. Amarapurkar, and S. Kankonkar, “Prevalence of hepatitis C genotypes in Indian patients and their clinical significance,” Journal of Association of Physicians of India, vol. 49, pp. 983–985, 2001.
[14]
S. Singh, V. Malhotra, and S. K. Sarin, “Distribution of hepatitis C virus genotypes in patients with chronic hepatitis C infection in India,” Indian Journal of Medical Research, vol. 119, no. 4, pp. 145–148, 2004.
[15]
I. Rehman, M. Idrees, M. Ali et al., “Hepatitis C virus genotype 3a with phylogenetically distinct origin is circulating in Pakistan,” Genetic Vaccines and Therapy, vol. 9, article 2, 2011.
[16]
H. Okamoto, Y. Sugiyama, S. Okada et al., “Typing hepatitis C virus by polymerase chain reaction with type-specific primers: Application to clinical surveys and tracing infectious sources,” Journal of General Virology, vol. 73, no. 3, pp. 673–679, 1992.
[17]
A. Ali, H. Ahmed, and M. Idrees, “Molecular epidemiology of Hepatitis C virus genotypes in Khyber Pakhtoonkhaw of Pakistan,” Virology Journal, vol. 7, pp. 203–210, 2010.
[18]
I. M. Idrees, H. Ahmed, M. Ali, L. Ali, and A. Ahmed, “Hepatitis C virus genotypes circulating in district Swat of Khyber Pakhtoonkhaw, Pakistan,” Virology Journal, vol. 8, article 16, 2011.
[19]
A. Z. Safi, Y. Waheed, J. Sadat, S. Salahuddin, U. Saeed, and M. Ashraf, “Molecular study of HCV detection, genotypes and their routes of transmission in North West Frontier Province, Pakistan,” Asian Pacific Journal of Tropical Biomedicine, vol. 2, no. 7, pp. 532–536, 2012.
[20]
S. Q. Afridi, M. N. Zahid, M. Z. Shabbir et al., “Prevalence of HCV genotypes in district Mardan,” Virology Journal, vol. 10, article 90, 2013.
[21]
T. Ohno, M. Mizokami, M. G. Saleh et al., “Usefulness and limitation of phylogenetic analysis for hepatitis C virus core region: Application to isolates from Egyptian and Yemeni patients,” Archives of Virology, vol. 141, no. 6, pp. 1101–1113, 1996.
[22]
G. L. Davis, R. Esteban-Mur, V. Rustgi et al., “Interferon alfa-2b alone or in combination with ribavirin for the treatment of relapse of chronic hepatitis C,” The New England Journal of Medicine, vol. 339, no. 21, pp. 1493–1499, 1998.
[23]
G. Dusheiko, J. Main, H. Thomas et al., “Ribavirin treatment for patients with chronic hepatitis C: results of a placebo-controlled study,” Journal of Hepatology, vol. 25, no. 5, pp. 591–598, 1996.
[24]
M. Idrees and S. Riazuddin, “Frequency distribution of hepatitis C virus genotypes in different geographical regions of Pakistan and their possible routes of transmission,” BMC Infectious Diseases, vol. 8, article 69, 2008.
[25]
S. Ali, I. Ali, S. Azam, and B. Ahmad, “Frequency distribution of HCV genotypes among chronic hepatitis C patients ofkhyber pakhtunkhwa,” Virology Journal, vol. 8, article 193, 2011.
[26]
M. Cenci, M. Massi, M. Alderisio, G. De Soccio, and O. Recchia, “Prevalence of hepatitis C virus (HCV) genotypes and increase of type 4 in Central Italy: an update and report of a new method of HCV genotyping,” Anticancer Research, vol. 27, no. 2, pp. 1219–1222, 2007.
[27]
A. Petruzziello, N. Coppola, A. M. Diodato et al., “Age and gender distribution of hepatitis C virus genotypes in the metropolitan area of Naples,” Intervirology, vol. 56, no. 3, pp. 206–212, 2013.
[28]
C. Henquell, C. Cartau, A. Abergel et al., “High prevalence of hepatitis C virus type 5 in central France evidenced by a prospective study from 1996 to 2002,” Journal of Clinical Microbiology, vol. 42, no. 7, pp. 3030–3035, 2004.
[29]
F. J. Sefidi, H. Keyvani, S. H. Monavari, S. M. Alavian, S. Fakhim, and F. Bokharaei-Salim, “Distribution of hepatitis C virus genotypes in Iranian chronic infected patients,” Hepatitis Monthly, vol. 13, no. 1, pp. e7991–e7998, 2013.
[30]
M. Martinot-Peignoux, F. Roudot-Thoraval, I. Mendel et al., “Hepatitis C virus genotypes in France: relationship with epidemiology, pathogenicity and response to interferon therapy,” Journal of Viral Hepatitis, vol. 6, no. 6, pp. 435–443, 1999.
[31]
Y. Waheed, T. Shafi, S. Z. Safi, and I. Qadri, “Hepatitis C virus in Pakistan: a systematic review of prevalence, genotypes and risk factors,” World Journal of Gastroenterology, vol. 15, no. 45, pp. 5647–5653, 2009.
[32]
W. Jafri, N. Jafri, J. Yakoob et al., “Hepatitis B and C: Prevalence and risk factors associated with seropositivity among children in Karachi, Pakistan,” BMC Infectious Diseases, vol. 6, article 101, 2006.
