Main Article Content

H.M Ahsan Sardar
Khalid Mahmood Anjum
Muhammad Imran
Usman Ahmad
Mohsin Munawar
Ahmed Imtanan Khalid
Farah Ejaz
Majid khan
Hamid ullah


Isothermal, Amplifications, LAMP, Diagnosing, Complications


Hepatitis C virus is considered a major reason of liver damaging, within estimated 180 million effected peoples worldwide. This study was conducted to diagnose the RNA of hepatitis C virus through isothermal amplification method (LAMP). Total genome of HCV different genotypes was aligned by MEGA CLUSTAL-W and different primers were designed by explorer version 4.0 software. All the extracted RNA of hepatitis C virus was collected from different laboratories located at different areas of Lahore, Pakistan. Extracted DNA of hepatitis B virus and other species was also included in this study to check the specificity of this technique. Persistence of hepatitis C virus was diagnosed by and isothermal amplification technique (LAMP) on dry heat block, incubator and thermo-cycler respectively. Ladder like pattern were obtained during this isothermal amplification for positive extracted RNA and no pattern was obtained for both positive and negative control samples in the population of Lahore, Pakistan. Significant difference was also observed in time consuming between LAMP and general PCR diagnosing technique. These results suggested that special measures should be adopted to diagnose the RNA of hepatitis C virus to reduce the incidence of complications of hepatitis C which increase the mortality among hepatitis C patients.

Abstract 60 | PDF Downloads 7


1. Gutierrez-Ruiz C, Robles-Diaz G, Kershenobich D. 2002. Emerging concepts in inflammation and fibrosis. Arc Med Res. 33: 595–599.
2. Sherlock S, Dooley J. 2002. Disease of the Liver and Biliary System. 11th ed., Blackwell Science Ltd, Oxford, UK, pp. 1-16.
3. Idrees M, Lal A, Naseem M, Khalid M. 2008. High prevalence of hepatitis C virus infection in the largest province of Pakistan. J Dig Dis. 9: 95-103.
4. Jehangir W, Ali F, Shahnawaz U, Iqbal T, Qureshi HJ. 2006. Prevalence of hepatitis B, C and HIV in blood donors of South Punjab. Esculapio. 2: 6-7.
5. Farhana M, Hussain I, Haroon TS. 2009. Hepatitis C: the dermatologic profile J Pak Assoc Derm. 18: 171-181.
6. Chaudhary IA, Samiullah U, Khan SS, Masood R, Sardar MA and Mallhi AA. 2007. Seroprevalence of hepatitis B and C among the healthy blood donors at Fauji Foundation Hospital, Rawalpindi. Pak J Med Sci. 23: 64-7.
7. Khan MSA, Khalid M, Ayub N, Javed M. 2004. Seroprevalence and risk factors of Hepatitis C virus (HCV) in Mardan, N.W.F.P. Rawal Med J. 29: 57-60.
8. Hashmie ZY, Chaudhary AH, Ahmad M, Ashraf M. 1999. Incidence of healthy voluntary blood donors at Faisalabad.Professional. J Pak Ass Derm 6: 551-5.
9. Kazmi Kato N. 2001. Molecular virology of hepatitis C virus. Acta Med Okayama. 55(3): 133-159.
10. Muhammad N, Jan A. 2005. Frequency of hepatitis C in Bunir, NWFP. J Coll Phys Surg Pak. 15: 11-14.
11. Tariq WU, Hussain AB, Karamat KA, Ghani E, Hussain T, Hussain S. 1999. Demographic aspects of hepatitis C in Northern Pakistan. J Pak Med Assoc. 49: 198-201.
12. El-Zayadi AR, Anis M. 2012. Hepatitis C virus induced insulin resistance impairs response to antiviral therapy. World J Gastroenterol. 18: 212-24.
13. Roberts AP, Lewis AP, Jopling CL. 2011. The role of microRNAs in viral infection. Prog Mol Biol Transl Sci. 102: 101-39.
14. Idrees M. 2008. Development of an improved genotyping assay for the detection of hepatitis C virus genotype and sub types in Pakistan. J Virol Method. 150: 50-56.
15. Notomi T, Okayama H, Masubuchi H, Yonekawa T, Watanabe K, Amino N. 2000. Loop-mediated isothermal amplification of DNA. Nucleic Acids Res. 12(28): E63-e63.
16. Yoneyama T, Kiyohara T, Shimasaki N, Kobayashi G, Ota Y, Notomi T, Totsuka A, Wakita T. 2007. Rapid and real-time detection of hepatitis A virus by reverse transcription loop-mediated isothermal amplification assay.J Virol Methods. 145(2): 162-8.
17. Lan X, Yang B, Li BY, Yin XP, Rui Li X, Liu JX. 2009. Reverse transcription loop-mediated isothermal amplification assay for rapid detection of hepatitis E virus. J Clin Microbiol 47: 2304–2306.
18. Moslemi E, Shahhosseiny M.H, Javadi G, Praivar K, Sattari TN, Amini HK. 2009. Loop mediated isothermal amplification (LAMP) for rapid detection of HBV in Iran. Afr J Microbiol Res. 3: 439–445.
19. Le Roux CA, Kubo T, Grobbelaar AA, Jansen Vuren P, Weyer J, Nel LH. 2009. Development and Evaluation of a Real-Time ReverseTranscription-Loop-Mediated Isothermal Amplification Assay for Rapid Detection of Rift Valley Fever Virus in Clinical Specimens. J Clin Microbiol. 4: 645-651.
20. Mori Y, Kitoa M, Tomita N, Notomi T. 2004. Real-time turbidimetryof LAMP reaction for quantifying template DNA . J Biochem Biophys Methods. 59: 45-157.
21. Mori Y, Nagamine K, Tomita N, Notomi T. 2001. Detection of loopmediated isothermal amplification reaction by turbidity derived from magnesium pyrophosphate formation. Biochem Biophys Res Commun. 289: 150-154.
22. Chen Hao-tai, Zhang Jie, Suna De-hui, Maa Li-na, Liu Xiang-tao, Quan Kai, Liu Yong-sheng 2008. Reverse transcription loopmediated isothermal amplification for the detection of highly pathogenic porcine reproductive and respiratory syndrome virus. J Virol Methods. 153: 266-268.
23. Hong TCT, Mai QL, Cuong DV, Parida MM, Harumi M, Tsugunori N, Hasebe F, and Morita K. 2004. Development and evaluation of a novel loop-mediated isothermal amplification method for rapid detection of severe acute respiratory syndrome coronavirus. J Clin Microbiol. 42: 1956-1961.
24. Parida MM, Guillermo P, Inoue S, Hasebe F, Morita K. 2004. RealTime Reverse Transcription Loop mediated isothermal amplification for rapid detection of West Nile Virus. J Clin Microbiol. 42: 257-263.
25. Pham HM, Nakajima C, Ohashi K, Onuma M. 2005. Loop-mediated isothermal amplification for rapid detection of Newcastle disease virus. J Clin Microbiol. 43: 1646-1650.
26. Poon LLM, Leung CSW, Chan KH, Lee JHC, Yuen KY, Guan Y. 2005. Detection of human influenza a viruses by loop-mediated isothermal Amplification. J Clin Microbiol. 43: 427-430.
27. Manns, M.P. 2001. Peginterferon alfa-2b plus ribavirin compared with interferon alfa-2b plus ribavirin for initial treatment of chronic hepatitis C: a randomised trial. Lancet. 358 (9286): p. 958-65.
28. Lakshmi V, Neeraja M, Lavanya V, Priyanka EN, Sharma S. 2016. Application of Real Time Loop Mediated Isothermal Amplification Assay on Dried Blood Spots in the Detection Of HCV RNA among High Risk Patients. J Emerg Dis Virol 2(1): doi
29. Kargar M, Askari A, Doosti A, and Ghorbani-Dalini S. 2012. Loop-Mediated Isothermal Amplification Assay for Rapid Detection of Hepatitis C virus. Indian J Virol. 23(1): 18–23. doi: 10.1007/s13337-012-0067-2.
30. Nyan DC, Swinson LK, 2016. A Method of Rapid Detection and Genotype identification of Hepatitis C Virus 1-6 by One Step Reverse Transcription loop-mediated isothermal amplification. Int J Inf Dis. 43: 30-36.
31. Gondeau Claire, Garcia Lydiane Pichard, Maurel Patrick. 2009. Cellular models for the screening and development of anti-hepatitis C virus agents. Pharmacol Ther. 124 (1): 1-22.
32. Pawlotsky JM. 2002. Use and interpretation of virological tests for hepatitis C. Hep. 36: 65–73. doi: 10.1002/hep.1840360709.
33. Lin BD, Evans MJ, Syder AJ, Wölk B, Tellinghuisen TL, Liu CC, Maruyama T, Hynes RO, Burton DR, McKeating JA, Rice CM. 2005. Complete replication of Hepatitis C virus in cell culture. Sci. 309: 623-626.
34. Vernet G. 2004. Molecular diagnostics in virology. J Clin Virol. 31: 239-247.
35. Antonelli A, Ferri C, Galeazzi M, Giannitti C, Manno D, Mieli-Vergani G 2008. HCV infection: pathogenesis, clinical manifestations and therapy. Clin Exp Rheumatol. 26 Suppl 48: S39-47.
36. El Serag HB. 2001. Epidemiology of hepatocellular carcinoma. Clin Liver Dis. 5: 87-107.
37. Chen LS, Morgan TR. 2006. The Natural History of Hepatitis C Virus (HCV) Infection. Int J Med Sci. 3(2): 47-52.
38. Wasley A, Alter MJ 2000. Epidemiology of hepatitis C virus :Geographic differences and temporal trends. Sem Liver Dis. 20: 1-16.
39. Perz Joseph F, Armstrong Gregory L, Farrington Leigh A, Hutin Yvan JF, Bell Beth P (2006a). The contributions of hepatitis B virus and hepatitis C virus infections to cirrhosis and primary liver cancer worldwide. J. Hepatol., 45(4): 529-538.
40. Perz Joseph F, Alter Miriam J (2006b).The coming wave of HCV-related liver disease:dilemmas and challenges. J Hepatol. 44: 441- 443.

Most read articles by the same author(s)

1 2 > >>