Journal of Isfahan Medical School

Journal of Isfahan Medical School

A Comparative Study of the Efficiency of the New Loop-Mediated Isothermal Amplification Assay and Routine Procedures for Diagnosis of Hepatitis C Virus

Document Type : Original Article (s)

Authors
Mohammad Kargar 1
Azam Askari 2
Abbas Doosti 3
Sadegh Ghorbani-Dalini 4
1 Associate Professor, Department of Microbiology, Islamic Azad University, Jahrom Branch, Jahrom, Iran
2 Department of Microbiology, Islamic Azad University, Jahrom Branch, Jahrom, Iran
3 Assistant Professor, Biotechnology Research Center, Islamic Azad University, Shahrekord Branch, Shahrekord, Iran
4 Department of Microbiology AND Young Researcher's Club, Islamic Azad University, Jahrom Branch, Jahrom, Iran
Abstract
Background: Hepatitis C virus (HCV) is a major cause of chronic liver disease and a public health problem. Therefore, rapid detection and management is important. This study aimed to compare the efficiency of enzyme-linked immunosorbent assay (ELISA), nested polymerase chain reaction (nested PCR), and loop mediated isothermal amplification assay (LAMP) for detection of HCV.Methods: This experimental study conducted on 50 serum samples suspected to HCV in Chaharmahal-va-Bakhtiari province, Iran. HCV Ab detected by ELISA and genomic RNA evaluated by nested PCR and LAMP techniques. Then, efficiency of ELISA and LAMP was calculated in comparison with nested PCR.Findings: Out of total samples, 45 (90%) samples were positive for HCV by nested PCR and LAMP. All samples (100%) were positive by ELISA, but 10% (5 samples) was false positive. In addition, in comparison with nested PCR, efficiency of LAMP and ELISA were 100% and 91%, respectively.Conclusion: Our results demonstrate that based on high efficiency, speed and cost effectiveness of LAMP assay, it could be used as a fast alternative method for diagnosis of HCV.
Keywords
Hepatitis C virus (HCV)
Enzyme-linked immunosorbent assay (Elisa)
Nested polymerase chain reaction (Nested-PCR)
Loop mediated isothermal amplification (LAMP)
  1. Brass V, Moradpour D, Blum HE. Molecular virology of hepatitis C virus (HCV): 2006 update. Int J Med Sci 2006; 3(2): 29-34.
  2. Sy T, Jamal MM. Epidemiology of hepatitis C virus (HCV) infection. Int J Med Sci 2006; 3(2): 41-6.
  3. Pawlotsky JM, Prescott L, Simmonds P, Pellet C, Laurent-Puig P, Labonne C, et al. Serological determination of hepatitis C virus genotype: comparison with a standardized genotyping assay. J Clin Microbiol 1997; 35(7): 1734-9.
  4. Pawlotsky JM. Molecular diagnosis of viral hepatitis. Gastroenterology 2002; 122(6): 1554-68.
  5. Pawlotsky JM, Bouvier-Alias M, Hezode C, Darthuy F, Remire J, Dhumeaux D. Standardization of hepatitis C virus RNA quantification. Hepatology 2000; 32(3): 654-9.
  6. Chevaliez S, Pawlotsky JM. Hepatitis C virus serologic and virologic tests and clinical diagnosis of HCV-related liver disease. Int J Med Sci 2006; 3(2): 35-40.
  7. Glynn SA, Wright DJ, Kleinman SH, Hirschkorn D, Tu Y, Heldebrant C, et al. Dynamics of viremia in early hepatitis C virus infection. Transfusion 2005; 45(6): 994-1002.
  8. Pawlotsky JM. Use and interpretation of virological tests for hepatitis C. Hepatology 2002; 36(5 Suppl 1): S65-S73.
  9. Wong T, Lee SS. Hepatitis C: a review for primary care physicians. CMAJ 2006; 174(5): 649-59.
  10. Notomi T, Okayama H, Masubuchi H, Yonekawa T, Watanabe K, Amino N, et al. Loop-mediated isothermal amplification of DNA. Nucleic Acids Res 2000; 28(12): E63.
  11. Askari A, Kargar M, Ghorbani-Dalini S, Doosti A. Loop-mediated isothermal amplification: Rapid and cost-effective method for detection of pathogens. Journal of Microbial World 2012; 5(1-2): 7-22. [In Persian].
  12. Parida M, Sannarangaiah S, Dash PK, Rao PV, Morita K. Loop mediated isothermal amplification (LAMP): a new generation of innovative gene amplification technique; perspectives in clinical diagnosis of infectious diseases. Rev Med Virol 2008; 18(6): 407-21.
  13. Mori N, Motegi Y, Shimamura Y, Ezaki T, Natsumeda T, Yonekawa T, et al. Development of a new method for diagnosis of rubella virus infection by reverse transcription-loop-mediated isothermal amplification. J Clin Microbiol 2006; 44(9): 3268-73.
  14. Ihira M, Sugiyama H, Enomoto Y, Higashimoto Y, Sugata K, Asano Y, et al. Direct detection of human herpesvirus 6 DNA in serum by variant specific loop-mediated isothermal amplification in hematopoietic stem cell transplant recipients. J Virol Methods 2010; 167(1): 103-6.
  15. Eiken Chemical Co. L. Eiken genome site [Online]. [cited 2005]; Available from: URL: http://loopamp.eiken.co.jp/e/lamp/
  16. Nagamine K, Hase T, Notomi T. Accelerated reaction by loop-mediated isothermal amplification using loop primers. Mol Cell Probes 2002; 16(3): 223-9.
  17. Kazemi B, Tafvizi F, Bandehpour M. Determination of HCV Genotypes in Iran. Biotechnology 2005; 4(2): 139-43.
  18. Kargar M, Askari A, Doosti A, Ghorbani-Dalini S. Loop-Mediated Isothermal Amplification Assay for Rapid Detection of Hepatitis C virus. Indian J Virol 2012; 23(1): 18-23.
  19. Kargar M, Baghernejad M, Doosti A. Compression of three methods of polymerase chain reaction, culture and rapid urease test in diagnosis of Helicobactr pylori in gastric biopsy specimen. Koomesh 2010; 11(3): 198-203. [In Persian].
  20. Swellam M, Mahmoud MS, Ali AA. Diagnosis of hepatitis C virus infection by enzyme-linked immunosorbent assay and reverse transcriptase-nested polymerase chain reaction: a comparative evaluation. IUBMB Life 2011; 63(6): 430-4.
  21. Parida MM. Rapid and real-time detection technologies for emerging viruses of biomedical importance. J Biosci 2008; 33(4): 617-28.
  22. Parida M, Horioke K, Ishida H, Dash PK, Saxena P, Jana AM, et al. Rapid detection and differentiation of dengue virus serotypes by a real-time reverse transcription-loop-mediated isothermal amplification assay. J Clin Microbiol 2005; 43(6): 2895-903.
  23. Mori Y, Nagamine K, Tomita N, Notomi T. Detection of loop-mediated isothermal amplification reaction by turbidity derived from magnesium pyrophosphate formation. Biochem Biophys Res Commun 2001; 289(1): 150-4.
  24. Parida MM, Santhosh SR, Dash PK, Tripathi NK, Saxena P, Ambuj S, et al. Development and evaluation of reverse transcription-loop-mediated isothermal amplification assay for rapid and real-time detection of Japanese encephalitis virus. J Clin Microbiol 2006; 44(11): 4172-8.
Journal of Isfahan Medical School
Volume 31, Issue 261 - Serial Number 261
September and October 2014
Pages 1876-1884

  • Receive Date 01 July 2013
  • Accept Date 06 April 2022