Comparison of RT-PCR and ELISA Methods in the Diagnosis of Hepatitis C Virus in Patients
,
Abstract
Background: Since HCV virus is the primary cause of liver cirrhosis and cancer, prompt diagnosis and timely treatment of this disease can prevent many complications. Due to the importance and necessity of this study, it aims to comparatively evaluate RT-PCR and ELISA methods in order to detect the presence of HCV infection in patients admitted to Baghiyatallah Hospital.
Methods: Sera of 49 patients admitted to Baghiyatallah Hospital since September, 2019 to August, 2020, were tested for RNA detection of HCV virus using RT-PCR and for the presence of anti-HCV antibody at the same time.
Results: In this study, the mean age of patients was approximately 38.3+6.3. The percentage of positive cases of HCV virus in the studied patients according to ELISA test and PCR test were 28.6% and 20.4%, respectively. Percentage of HCV positive cases had p = 0.001 based on PCR and ELISA tests by age group, number of sexual partners, history previous HCV infection, liver cirrhosis, addiction and treatment of previous HCV infection which was significantly different; However, it was not significant in terms of gender (p = 0.5232). According to Kappa coefficient, the percentage of agreement is 91.8 in both methods which indicates that the two models are consistent (P = 0.001) and the diagnostic value of ELISA versus PCR with sensitivity and specificity were 100% and 89.7%, respectively.
Conclusion: ELISA susceptibility to anti-HCV antibody is more than 99%, but its specificity is low compared to RT-PCR method. On the other hand, these tests show only hepatitis C affection and does not distinguish between chronic acute or improved infection. It is not able to detect patients in the window phase, so the simultaneous application of ELISA and molecular methods is recommended to diagnose and follow the treatment of HCV virus.
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22. Narayan Singh M, Arjun Lal, Kumar Poddar C, et al. Comparative evaluation of ELISA and rapid screening techniques for the diagnosis of HCV. JEMDS 2017; 6(93):6683-8.
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28. Rasheed N, Abdullah Balatay A , Turan A. The distribution of HCV in subjects attending hospitals in Duhok City, Iraq. Asian Pac J Trop Biomed 2017; 7(3):262-4.
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31. Francois Rouet, Deleplancque LVC, Mboumba BB, et al. Usefulness of a fourth generation ELISA assay for the reliable identification of infection PlosOne 2015; 10(1):116975.
32. Ahmed SHH, Ibrahim AM, Abo-El-Azaem NGM, et al. Evaluation of some available HCV antibody detection tests ELISA chemi-luminescence Immune Assay and RT.PCR assay in the diagnosis of Hepatitis C virus infection. EJHM 2018; 72(7):4874-9 .
33. Taheri-Ghahfarkhi F, Tajbakhsh E, Heydari-Soureshjani E. Molecular detection if HCV and HBV in HIV positive patients in Chaharmahal and Bakhtiari province. Sci J Iran Blood Transfus Organ 2018: 15(4):301-9.
34. Khazaee T, Ebrahimzadeh A, Moghaddam E. Assessment of prevalence and Determine Infections of Hepatitis c and Hepatitis D in patients with chronic Hepatitis B. Birjand Uni Med Sci 2016; 20:230-6.
35. Garg G, Kumar D, Asim M, et al. Multiplex Reverse Transcriptase-PCR for simultaneous detection of hepatitis B, C, and E viruses. Department of Medicine University of Dehli 2016; 33:39.
36. Omar MZ, Metwally MA, El-Feky HM, et al. Role of intra familial transmission in high prevalence of hepatitis C virus in Egypt. Department of Hepatology 2017; 36:58-63.
37. You MW, Kim KW, Shim J, et al. Impact of liver stiffness measurement on hepatocellular carcinoma development in chronic hepatitis C patients. J Gastroenterol Hepatol 2021; 36(3):601-8 .
38. Nyberg AH, Sadikova E, Cheetham C, et al. Increased cancer rates in patients with chronic hepatitis C. Liver international 2020; 40(3):685-93.
39. Minola E, Prati D, Suter F, et al. Age at infection affects the long term outcome of transfusion associated chronic hepatitis c. American J Hematology 2002; 99(12):4588-91.
40. Hedayati-Moghaddam MR, Soltanian H, Ahmadi S, et al. Occult hepatitis c virus infection in the Middle East and Eastern Mediterranean countries; a systematic review and meta-analysis. World J Hepatology 2021; 13(2):242.
41. Azza Galal farghaly, Yasmine Mohammed, Alkassabany Engy Mohamed , El-Ghitany. HBV-HCV and HIV among famele sex workers. SRT 2020; 35(4):462-77.
42. Kakchapati S, Bir MM, Rawal B, et al. Social determinants and risk behaviors associated with prevalent Hepatitis c and HIV/HCV co infection among male injection drug users in Nepal. Archives of public Health 2017; 75(1):1-10.
2. Mahmud S, Akbarzade V, Abu-Radded Lj. The epidemiology of hepatitis C virus in Iran:systematic review and meta-analyses Sci Rep 2018; 8(1):150.
3. Taherkhani R, Farshadpour F. Epidemiology of hepatitis C virus in Iran, Word J Gastroenterol 2015; 21(38):10790-810.
4. Taherkhani R, Farshadpour F. Global elimination of hepatitis virus infection: progresses and the remaining challenges. World J Hepatol 2017; 9(33):1239-52.
5. Bahadar N, Khan F, Israr M, et al. The correlation between RT-PCR and Elisa assay on hepatitis C positive serum samples. Pure Appl Bi 2016; 25:87-100.
6. El-Shamy A, Hottah Impact of hepatitis C virus heterogenity on interferon sesitivity an overview. World J Gastroentreol 2014; 20: 7555-756.
7. Aisyah DN, Shall Cross L, Hully AJ, et al. Assessing hepatitis C spontaneous clearance and understanding associated factors -A systematic review and meta-analysis. J Viral Hepat 2018; 25:680-698.
8. WHO,Global health sector strategy on viral hepatitis 2016-2021. Towards ending viral hepatitis. 2021.
9. Liu HY, Hopping GC, Vaidyanathan U, et al. Polymerase chain reaction and its application in the diagnosis of infectious keratitis. Med Hypothesis Discov Innov Ophthalmol 2019; 8(3):152-155.
10. Laperche S1, le Marrenc N, Givault A, et al. Simultaneous detection of hepatitis c virus core antigen and anti hcv antibodies improves the early detection of hcv infection. J Clin Microbial 2005; 43(8):3877-83.
11. Lambert N. value of hcv antigen antibody combined hcv assay in hepatitis c diagnosis. Dev Biol 2007; 137 :113-21.
12. Shen C, Jiang X, Li M, et al. Hepatitis Virus and Hepatocellular Carcinoma: Rec Adv Cancers (Basel) 2023; 15(2):533.
13. World Health Organization. Global hepatitis report, 2017.
14. Stasi C, Silvestri C, Voller F. Update on Hepatitis C Epidemiology: Unaware and Untreated. SN Compre Clin Med 2020; 2:2808-15.
15. Armstrong GL, Wasley A, Simard EP, et al. The prevalence of hepatitis virus infection in the United States 1999 through 2002; 144:705-14.
16. Khodabandehloo M, Roshani D. Prevalence of hepatitis C virus genotypes in Iranian patients: a systematic review and meta-analysis. Hepat Mon 2014; 14(12):e22915.
17. Bourliere M, penarada G, Khiri H, et al. Real time PCR assays for hepatitis C virus RNA quantitation are adequate for clinical management of chronic HCV infection. J clin Microbiol 2006: 44;2507-11.
18. Wei Liu, Xiwen Jiang, Yue Liu, et al. Bioinformatics analysis of quantitative PCR and reverse teanscription PCR. Curr Bioinformatics 2019; 14(5):400.
19. Ullah S, Sohail Ahmad S, Qaisar A, et al. Current screening strategy poses risk of spreading of hepatitis C virus infection. Int J Front Sci 2020; 5(1):256-61.
20. Peng Y, Zhenrui X, Tlaoyang T, et al. Fast and ultrasensitivity ELISA Rolling circle Amplification 2021; 25:236-241.
21. Hosiny Ahmed H, Mohamad A, Ibrahim N, et al. Evaluation of some available HCV antibody detection tests (ELISA. RT.PCR) assay in the diagnosis of hepatitis c virus infection. EJHM 2018; 72(7):4874-9.
22. Narayan Singh M, Arjun Lal, Kumar Poddar C, et al. Comparative evaluation of ELISA and rapid screening techniques for the diagnosis of HCV. JEMDS 2017; 6(93):6683-8.
23. Mohammadi S, Khodabandehloo M. Prevalence of Hepatitis C Virus Antibodies among Beta-Thalassemia Major Patients in Kurdistan Province, Iran. Arch Clin Infect Dis 2017; 12(3):e62419.
24. Ashrath WS, Keum-Soo S, Dilipkumar P, et al. Developments in the HCV screening technologies based on the detection of antigens and antibodies. Sensors (Basel, Switzerland). 2019; 19:4257.
25. Rujipat W, Preeyaporn V, Chompoonut A, et al. HCV core antigen is an alternative marker to HCV RNA for evaluating active HCV infection: implications for improved diagnostic option in an era of affordable DAAs. PeerJ 2017; 5:e4008.
26. Arıdoğan BC, Aynali A, Kaya S, et al. Comparison of HCV core antigen and anti-HCV with HCV RNA results. African Health Sci 2015; 14(4):816.
27. Samimi-Rad K, Shahbaz B. Hepatitis C virus genotypes among patients with thalassemia and inherited bleeding disorders in Markazi province, Iran. Haemophilia 2007; 13(2):156-63.
28. Rasheed N, Abdullah Balatay A , Turan A. The distribution of HCV in subjects attending hospitals in Duhok City, Iraq. Asian Pac J Trop Biomed 2017; 7(3):262-4.
29. El-Sokkary R, Tash E, Meawed E. Detection of hepatitis C virus (HCV) among health care providers in an Egyptian university hospital: different diagnostic modalities. Infection and Drug Resistance 2017:10;357-64.
30. Muhammad umer khan, Haleema sadia, Asma Irshad. Baing A.A. Ashiq S, Zabid B, et al. Detection quantification and genotype distribution of hcv paticentsion laahore,pakistan by real-time PCR. Afri Health 2020; 20(3):1143-51.
31. Francois Rouet, Deleplancque LVC, Mboumba BB, et al. Usefulness of a fourth generation ELISA assay for the reliable identification of infection PlosOne 2015; 10(1):116975.
32. Ahmed SHH, Ibrahim AM, Abo-El-Azaem NGM, et al. Evaluation of some available HCV antibody detection tests ELISA chemi-luminescence Immune Assay and RT.PCR assay in the diagnosis of Hepatitis C virus infection. EJHM 2018; 72(7):4874-9 .
33. Taheri-Ghahfarkhi F, Tajbakhsh E, Heydari-Soureshjani E. Molecular detection if HCV and HBV in HIV positive patients in Chaharmahal and Bakhtiari province. Sci J Iran Blood Transfus Organ 2018: 15(4):301-9.
34. Khazaee T, Ebrahimzadeh A, Moghaddam E. Assessment of prevalence and Determine Infections of Hepatitis c and Hepatitis D in patients with chronic Hepatitis B. Birjand Uni Med Sci 2016; 20:230-6.
35. Garg G, Kumar D, Asim M, et al. Multiplex Reverse Transcriptase-PCR for simultaneous detection of hepatitis B, C, and E viruses. Department of Medicine University of Dehli 2016; 33:39.
36. Omar MZ, Metwally MA, El-Feky HM, et al. Role of intra familial transmission in high prevalence of hepatitis C virus in Egypt. Department of Hepatology 2017; 36:58-63.
37. You MW, Kim KW, Shim J, et al. Impact of liver stiffness measurement on hepatocellular carcinoma development in chronic hepatitis C patients. J Gastroenterol Hepatol 2021; 36(3):601-8 .
38. Nyberg AH, Sadikova E, Cheetham C, et al. Increased cancer rates in patients with chronic hepatitis C. Liver international 2020; 40(3):685-93.
39. Minola E, Prati D, Suter F, et al. Age at infection affects the long term outcome of transfusion associated chronic hepatitis c. American J Hematology 2002; 99(12):4588-91.
40. Hedayati-Moghaddam MR, Soltanian H, Ahmadi S, et al. Occult hepatitis c virus infection in the Middle East and Eastern Mediterranean countries; a systematic review and meta-analysis. World J Hepatology 2021; 13(2):242.
41. Azza Galal farghaly, Yasmine Mohammed, Alkassabany Engy Mohamed , El-Ghitany. HBV-HCV and HIV among famele sex workers. SRT 2020; 35(4):462-77.
42. Kakchapati S, Bir MM, Rawal B, et al. Social determinants and risk behaviors associated with prevalent Hepatitis c and HIV/HCV co infection among male injection drug users in Nepal. Archives of public Health 2017; 75(1):1-10.
| Files | ||
| Issue | Vol 12 No 2 (2024) | |
| Section | Original Articles | |
| DOI | https://doi.org/10.18502/jmb.v12i2.15623 | |
| Keywords | ||
| Hepatitis C Viruses RT- PCR Enzyme-Linked Immunosorbent Assay | ||
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How to Cite
1.
Beik Mahdavi F, Bashizadehfakhar H, Jalalian M, Rangraz S. Comparison of RT-PCR and ELISA Methods in the Diagnosis of Hepatitis C Virus in Patients. J Med Bacteriol. 2024;12(2):34-42.
