Detection of Hepatitis C Virus RNA in Blood Donors with Nested PCR Technique

Supiana Dian Nurtjahyani, Mochammad Amin Amin, Retno Handajani Handajani


Objective: This study was conducted to detect hepatitis C virus (HCV) RNA among blood donors in Tuban, East Java, Indonesia,  using nested Polymerase Chain Reaction (PCR) technique.

Material and methods: Seven blood donors  with positives anti-HCV were obtained from 500 blood donors from Red Cross Blood Donor Center (Palang Merah Indonesia = PMI) Tuban, East Java, Indonesia, consisting of 375 men (75%) and 125 women (25%). Detection of anti-HCV antibody prevalence in the blood donors was done by using Enzyme-Linked Immunosorbent Assay (ELISA).  The detection of HCV RNA in blood donors  sera  with positives  anti-HCV were performed using nested-PCR technique with three set of primer pairs targeting the NS5B and 5’UTR regions of HCV.  Further amplification products were visualized on a 2% agarose gel containing ethidium bromide under  UV illumination.

Result: Seven positives anti-HCV blood donors sera (1.4%) were obtained from 500 blood donors sera, in PMI Tuban, East Java, Indonesia. Out of seven blood donors sera with positives anti-HCV, 6 sera (85,7%)  were positives HCV RNA by using nested PCR technique. Five HCV RNAs were detected with two set of primers based on NS5B HCV region and 1 positive HCV RNA was detected with another set of primer based on 5’UTR HCV region.

Conclusion: HCV RNAs were detected in 85,7% positives anti-HCV blood donors using nested PCR technique, in PMI Tuban, East Java. Further research is needed to determine the  genotype  and  subtype  of  HCV.



Anti-hepatitis C virus; Virus Hepatitis C; Nested-PCR; Blood donors Indonesia

Full Text:



World Health Organization. 2002. Hepatitis C. WHO/CDS/CSR/)LYO/2003

Sharma SD. 2010. Hepatitis C Virus: Molecular Biology Current Therapeutic Options. Indian J Med Res. 131:17-34.

Lavanchy D (2011). Evolving Epidemiology ofHepatitis C Virus. Clin Mcrobiol Infect, 17: 107.

Friedman LS. 2013. Liver, Biliary Tract, & Pancreas Disorders. In: Current Medical Diagnosis & Treatment. Fifty- Second Edition. Editors: Papadakis MA, McPhee SJA Rabow MW. New York, pp.662-682.

Simmonds P, Bukh J, Combet C, Deleage G, Enomoto N. 2005. Consensus Proposals for a Unified System of Nomenclature of Hepatiti C Virus Genotypes. Hepatology. 42: 962.

Murphy, D. G., Willems, B., Deschenes, M., Hilzenrat, N., Mousseau, R. & Sabbah, S. 2007. Use of Sequence Analysis of the NS5B Region for outine Genotyping of Hepatitis C Virus with Reference to C/E1 and 5’Untranslated Region Sequences. J Clin.Microbiol 45,1102-112.

Allison RD, Camtilena CC, Koziol D, Schechterly C, Ness P, Gihble J, Kleiner DE, Ghany MG, Alter HJ. 2012. A 25-Year Study of the Clinical and Histologic Outcomes of Hepatitis C Virus Infection and Its Modes of Transmission in a Cohort of Initially Asymptomatic Blood Donors. The Journal of Infectious Diseases, 206: 654.

Lina MR, Bela B, Dadang S. 2004. Uji PCR (Polymerase Chain Reaction ) untuk Virus Hepatitis C. Risalah Seminar Ilmiah Penelitian dan Pengembangan Aplikasi Isotop dan Radiasi.

Kemenkes RI 2014. Infodatin: Pusat Data dan Informasi Kementrian Kesehatan RI: Situasi dan Analisis Hepatitis. Jakarta.

Soetjipto, Handajani R, Lusida MI, Darmadi S, Adi P, Soemarto, Ishido S, Katayama Y, Hotta H, 1996. Differential Prevalence of Hepatitis C Virus Subtypes i healthy blood Donors, Patients on Maintenace Hemodialysis, and Patients with Hepatocellular Carcinoma in Surabaya, Indonesia. J. Clin. Microbiol. 34(12): 2875-2880.

Anggorowati N, YoshihikoYano,Didik Setyo Heriyanto, Hanggoro Tri Rinonce, Takako Utsumi, Deshinta Putri Mulya, et al. 2012.Clinical and Virological Characteristics of Hepatitis B or C Virus Co-Infection With HIV in Indonesian Patients. J.Med.Virol.84:857-65.

Mori, S., N. Kato, A. Yagyu, T. Tanaka, Y. Ikeda, B. Petchlai, P. Chiewsilp, T. Kurimura, and K. Shimotohno. 1992. A new type of hepatitis C virus in patients in Thailand. Biochem. Biophys. Res. Commun. 183:334-342.

Apichartpiyakul C, Chittivudikarn C, Miyajima H, Homma M, Hotta H. 1994. Analysis of Hepatitis C Virus Isolates among Healthy Blood Donors and Drug Addicts in Chiang Mai,Thailand.Journal of Clinical Microbiology. 32(9): 2276-2279.

Doi H, Apichartpiyakul C, Ohba KI, Mizokami M and Hotta H. 1996. Hepatitis C Virus (HCV) Subtype Prevalence in Chiang Mai, Thailand, and Identification of Novel Subtypes Of HCV Major Type 6. J Clin Microbiol 34: 569 574.

Nakatani SM. Santos CA, Riediger IN, Krieger MA, Duarte CAB, Debur MdD, Carrilho FJ, Ono SK. 2011. Comparative Performance Evaluation of Hepatitis C Virus Genotyping Based on the 5’ Untranslated Region Versus Partial Sequencing of the NS5B Region of Brazilian Patients with Chronic Hepatitis C.Virology Journal. 8: 459.

Marascio N, Torti C, Liberto MC, Foca A. 2014. Update on Different Aspects of HCV Variablity: Focus on NS5B Polymerase. BMC Infectious Diseases. 14:1471-2334.

Juniastuti, Takako Utsumi, Nasronudin, Lindawati Alim Sardjono, Mochamad Amin, Myrna Adianti, Yoshiiko Yano, Soetjipto, Yoshitake Hayashi and Hak Hotta. 2014 High Rate of Seronegative HCV Infection In HIV-Positive Patients. Biomedical Reports. 2: 79-84.

Suwarso, 1993. Virus Hepatitis-C. Berkala Ilmu Kedokteran. XXV (3): 131-137


  • There are currently no refbacks.