Skip to content
Research Article Open access CC BY 4.0

Molecular Detection of Hepatitis C Virus (HCV) by Conventional One-step RT-PCR Coupled with Nested PCR

Vikrant Sharma, Deepak Kumar, Divya Dhull, Sulochana Kaushik, Jaya Parkash Yadav, Samander Kaushik

International Blood Research & Reviews · pp. 1–6 · Published 15 Jun 2017

10.9734/IBRR/2017/34463

Abstract

Aims: HCV causes both acute and chronic infections and can be easily transmitted through contaminated blood or other body fluids. The present study deals with the molecular detection of HCV with help of one-step RT-PCR assay followed by nested PCR and agarose gel electrophoresis.  Study Design: RNA extracted from the confirmed positive samples of HCV was utilized for the standardization of the one-step RT-PCR assay and nested PCR assay for diagnosis of HCV. Place and Duration of Study: Centre for Biotechnology, Maharshi Dayanand University, Rohtak Haryana, India, during period of one year (January-December 2015). Methodology: HCV positive samples were obtained from Department of Medicine, Maulana Azad Medical College (MAMC), New Delhi, India. Published primers from most conserved regions of HCV were taken and these primers were able to amplify all the strains of HCV. One-step RT-PCR kits, primers, extracted RNA from these positive samples were used for standardization of molecular diagnostic assays. The results were checked by 2% agarose gel electrophoresis. Results: Positive samples of HCV were detected by nested PCR. Positive samples showed sharp band of 405bp while there was no amplification in the negative control. Conclusion: Rapid tests have low sensitivity and specificity while molecular assays are rapid, sensitive and specific. Conventional one-step RT-PCR assay followed by nested PCR is rapid, specific, sensitive and it is also less costly than real-time RT-PCR. Cost of an assay is an important factor in controlling a disease in resource limited settings of developing countries.  

Hepatocellular carcinoma conventional PCR viral hepatitis hepatitis C virus one-step RT-PCR, nested PCR, molecular diagnosis

References (21)

  1. 1 Isolation of a cDNA cLone Derived from a Blood-Borne Non-A, Non-B Viral Hepatitis Genome [DOI]
  2. 2 Global epidemiology of hepatitis C virus infection: New estimates of age-specific antibody to HCV seroprevalence [DOI]
  3. 3 Harrison's Principles of Internal Medicine. [DOI]
  4. 4 Consensus Proposals for a Unified System of Nomenclature of Hepatitis C Virus Genotypes * [DOI]
  5. 5 Evolving epidemiology of hepatitis C virus [DOI]
  6. 6 Internal ribosome entry site within hepatitis C virus RNA [DOI]
  7. 7 Use and interpretation of virological tests for hepatitis C [DOI]
  8. 8 Hepatitis C: a review for primary care physicians [DOI]
  9. 9 Diagnosis and treatment of chronic hepatitis C infection [DOI]
  10. 10 Real-Time PCR Assays for Hepatitis C Virus (HCV) RNA Quantitation Are Adequate for Clinical Management of Patients with Chronic HCV Infection [DOI]
  11. 11 Community-based Study of Hepatitis C Virus Infection and Type 2 Diabetes: An Association Affected by Age and Hepatitis Severity Status [DOI]
  12. 12 Analysis of the 5′ Noncoding Region versus the NS5b Region in Genotyping Hepatitis C Virus Isolates from Blood Donors in France [DOI]
  13. 13 Diagnosis of influenza viruses with special reference to novel H1N1 2009 influenza virus [DOI]
  14. 14 Comparison of multiplex RT-PCR with virus isolation for detection, typing and sub-typing of influenza virus from influenza-like illness cases [DOI]
  15. 15 Acquisition of susceptibility to hepatitis C virus replication in HepG2 cells by fusion with primary human hepatocytes: Establishment of a quantitative assay for hepatitis C virus infectivity in a cell culture system [DOI]
  16. 16 Nested Restriction Site-Specific PCR To Detect and Type Hepatitis C Virus (HCV): a Rapid Method To Distinguish HCV Subtype 1b from Other Genotypes [DOI]
  17. 17 Influence of quasispecies on virological responses and disease severity in patients with chronic hepatitis C [DOI]
  18. 18 Comparative Analysis of Molecular Methods for Detection of Influenza Viruses [DOI]
  19. 19 Evaluation of PCR and nested PCR for laboratory diagnosis of hepatitis C virus infection [DOI]
  20. 20 Detection of hepatitis C virus RNA by a combined reverse transcription PCR assay: comparison with nested amplification and antibody testing [DOI]
  21. 21 Molecular Detection of Hepatitis B Viruses (HBV) [DOI]

Cited by 1

Study of EBV and Associated Gastric Malignancies

B. Supraja, Vardarajalu, P. Karkuzhali · 2021

Related research

Article metrics

Real usage data collected on this platform.

40

Page views

0

PDF downloads

6

Outbound clicks

1

Citations

Views over time

Views by country

Approximate, from request IP at view time — not citizenship or institution. Countries with fewer than 5 views are grouped as "Other".

Traffic sources

Referring site, by host.

Views and downloads exclude known bots/crawlers. Citations combines this platform's own DOI-resolved index with each external source's own reported total — see Cited by above for individually listed citing works. Last refreshed 0 seconds ago.