Determination of anti-HCV signal to cut-off value in patients with hepatitis C virus infection and the variety of antibody responses

Year 2023, Volume: 9 Issue: 3, 484 - 494, 04.05.2023

Murat Ocal Mehmet Emin Bulut

https://doi.org/10.18621/eurj.945588

Abstract

Objectives: The diagnosis of hepatitis C virus (HCV) infection starts with the detection of antibodies against recombinant or synthetic HCV proteins by Enzyme Immunoassay (EIA). Although EIA tests are highly sensitive, false positivity rates are not low. Positive anti-HCV results are generally confirmed with complementary tests such as Nucleic Acid Amplification Tests (NAAT), or Western Blot modifications.

Methods: The anti-HCV results of 199,516 individuals referred from various clinics between 2015 and 2019 were evaluated retrospectively at University of Health Sciences, Şişli Hamidiye Etfal Training and Research Hospital, Medical Microbiology Laboratory. From the 2039 samples, of which EIA tests resulted borderline and reactive, 1419 samples having Line Immunoassay (LIA) confirmatory test results were included in the study.

Results: LIA tests yielded positive, negative and indeterminate for 820 (57.8%), 519 (36.6%) and 80 (5.6%) of 1419 samples, respectively. The optimal threshold point for EIA anti-HCV signal to cut-off (S/Co) according to LIA was found to be 15.85 corresponded to diagnostic sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) and accuracy of 94.9%, 94.8%, 96.6%, 92.1%, 94.9%, respectively. The most common proteins detected in LIA positive samples were C1 96.3%, C2 90.4%, and NS3 93.2%.

Conclusions: To prevent false positivities, confirmatory tests must be used for samples with low S/Co ratios. The use of S/Co value will make significant contribution to reducing both false-positive results and the LIA confirmatory test consumption. There was no correlation between the number of bands and EIA index values in LIA positive samples, while the relationship between the number of 3+ bands and index values was remarkable.

Keywords

Hepatitis C virus, signal to cut-off, enzyme immunoassay, line immunoassay, confirmatory test

References

• 1. Selek MB, Baylan O, Karagöz E, Özyurt M. Changes in hepatitis C virus genotype distribution in chronic hepatitis C infection patients. Indian J Med Microbiol 2018;36:416-21.
• 2. Ergünay K, Abacıoğlu H. [Clinical impact of hepatitis C virus genomic variations]. Mikrobiyol Bul 2015; 49(4): 625-635. [Article in Turkish]
• 3. Arora S, Doda V. Role of signal-to-cut-off ratios of anti-hepatitis C virus antibody by enzyme immunoassays along with ID-NAT for screening of whole blood donors in India. Asian J Transfus Sci 2016;10:75-8.
• 4. Tabak F. Enfeksiyon Hastalıkları, 4th.ed., İstanbul Tıp Kitapevi: Istanbul, 2019.
• 5. Tiryaki Y, Duran AC, Ozcolpan OO. Distribution of hepatitis c virus genotypes in Aydın province. Viral Hepat J 2018;24:70-4.
• 6. Petruzziello A, Marigliano S, Loquercio G, Cozzolino A, Cacciapuoti C. Global epidemiology of hepatitis C virus infection: an up-date of the distribution and circulation of hepatitis C virus genotypes. World J Gastroenterol 2016;22:7824.
• 7. Mazzarella C, Rocco C, Vallefuoco L, Sorrentino R, Braschi U, Lauritano G, et al. Differential reactivity of anti-hepatitis C virus screening assays in patients with waning antibodies. Future Virol 2019;14:303-9.
• 8. Choi MS, Lee K, Hong YJ, Song EY, Kim DS, Song J. The role of the signal-to-cutoff ratio in automated Anti-HCV chemiluminescent immunoassays by referring to the nucleic acid amplification test and the recombinant immunoblot assay. Ann Lab Med 2018;38:466-72.
• 9. Getchell JP, Wroblewski KE, DeMaria Jr A, Bean C L, Parker MM, Pandori M, et al. Testing for HCV infection: an update of guidance for clinicians and laboratorians. MMWR. Morb Mortal Wkly Rep 2013;62: 362-5.
• 10. Rafik M, Bakr S, Soliman D, Mohammed N, Ragab D, Abd ElHady W, Samir N. Characterization of differential antibody production against hepatitis C virus in different HCV infection status. Virol J 2016;13:116.
• 11. Tsukiyama-Kohara K, Kohara M. Hepatitis C virus: viral quasispecies and genotypes. Int J Mol Sci 2018;19:23.
• 12. Altuğlu I, Gürsel D, Aksoy A, Orman M, Erensoy S. The importance and role of anti-HCV signal/cutoff ratio in diagnosis of hepatitis C virus infection. Ege J Med 2011;50:223-8.
• 13. Warkad SD, Song KS, Pal D, Nimse S. B. Developments in the HCV screening technologies based on the detection of antigens and antibodies. Sensors (Basel) 2019;19:4257.
• 14. Sirin MC, Aridogan BC, Cetin ES, Sirin FB. Evaluation of biochemical, hematological, RIBA and PCR assays in predicting viremia in anti-HCV positive patients. J Infect Dev Ctries 2019;13:736-43.
• 15. Erensoy S. Diagnosis of hepatitis C virus (HCV) infection and laboratory monitoring of its therapy. J Clin Virol 2001;21:271-81.
• 16. Alter MJ, Kuhnert WL, Finelli L. Centers for Disease Control and Prevention. Guidelines for laboratory testing and result reporting of antibody to hepatitis C virus. Centers for Disease Control and Prevention. MMWR Recomm Rep 2003;52:1-13, 5; quiz CE1-4.
• 17. Zhang K, Wang L, Lin G, Li J. Is anti-hepatitis C virus antibody level an appropriate marker to preclude the need for supplemental testing? Intervirology 2015;58:310-7.
• 18. Kodani M, Martin M, de Castro VL, Drobeniuc J, Kamili S. An automated immunoblot method for detection of IgG antibodies to hepatitis C virus: a potential supplemental antibody confirmatory assay. J Clin Microbiol 2019;57:e1567-18.
• 19. Seo YS, Jung ES, Kim JH, Jung YK, Kim JH, An H, et al. Significance of anti-HCV signal-to-cutoff ratio in predicting hepatitis C viremia. Korean J Intern Med 2009;24:302-8.
• 20. Gupta E, Bajpai M, Choudhary A. Hepatitis C virus: screening, diagnosis, and interpretation of laboratory assays. Asian J Transfus Sci 2014;8:19-25.
• 21. Kalem F, Yüksekkaya Ş, Türk Dagi H, Ertuğrul Ö, Doğan M. Comparative evaluation of automated chemiluminescence tests and RIBA assay used in HCV diagnosis. Biomed Res J 2016;27:1261-4.
• 22. Centers for Disease Control and Prevention. Hepatitis C. Available at: https://www.cdc.gov/hepatitis/hcv/labtesting.htm (12.12.2017).
• 23. Aydin G, Adaleti R, Boz ES, Yücel FM, Özhan HK, Aksaray S. [Investigation of anti-HCV S/CO value in detecting viremia in patients with hepatitis C virus infection]. Mikrobiyol Bul 2020; 54:110-9. [Article in Turkish]
• 24. Karakoc AE, Berkem R, Irmak H, Demiroz AP, Yenicesu I, Ertugrul N, et al. Investigation of an algorithm for anti HCV EIA reactivity in blood donor screening in Turkey in the absence of nucleic acid amplification screening. Transfus Apher Sci 2017;56:732-7.
• 25. Pan J, Li X, He G, Yuan S, Feng P, Zhang X. Reflex threshold of signal-to-cut-off ratios of the Elecsys anti-HCV II assay for hepatitis C virus infection. J Infect Dev Ctries 2016;10:1031-4.
• 26. Kao HH, Chen KS, Lin CL, Chang JJ, Lee CH. Utilization of signal-to-cutoff ratio of hepatitis C virus antibody assay in predicting HCV viremia among hemodialysis patients. Nephron 2015;130:127-33.
• 27. Makuria AT, Raghuraman S, Burbelo PD, Cantilena CC, Allison RD, Gibble J, et al. The clinical relevance of persistent recombinant immunoblot assay–indeterminate reactions: insights into the natural history of hepatitis C virus infection and implications for donor counseling. Transfusion 2012;52:1940-8.
• 28. Hitziger T, Schmidt M, Schottstedt V, Hennig H, Schumann A, Ross S, et al. Cellular immune response to hepatitis C virus (HCV) in nonviremic blood donors with indeterminate anti-HCV reactivity. Transfusion 2009;49:1306-13.
• 29. Seeff LB, Hollinger FB, Alter HJ, Wright EC, Cain CM, Buskell Z, et al. Long-term mortality and morbidity of transfusion-associated non-A, non-B, and type C hepatitis: a National Heart, Lung, and Blood Institute collaborative study. Hepatology 2001;33:455-63.
• 30. Kondili LA, Chionne P, Costantino A, Villano U, Noce CL, Pannozzo F, et al. Infection rate and spontaneous seroreversion of anti-hepatitis C virus during the natural course of hepatitis C virus infection in the general population. Gut 2002;50:693-6.
• 31. Pereira FM, Zarife MAS, Reis EAG, Reis MG. Indeterminate RIBA results were associated with the absence of hepatitis C virus RNA (HCV-RNA) in blood donors. Rev Soc Bras Med Trop 2014;47:12-7.
• 32. Kittlesen DJ, Chianese-Bullock KA, Yao ZQ, Braciale TJ, Hahn YS. Interaction between complement receptor gC1qR and hepatitis C virus core protein inhibits T-lymphocyte proliferation. J Clin Invest 2000;106:1239-49.
• 33. Beld M, Penning M, van Putten M, Lukashov V, van den Hoek A, McMorrow M, et al. Quantitative antibody responses to structural (Core) and nonstructural (NS3, NS4, and NS5) hepatitis C virus proteins among seroconverting injecting drug users: impact of epitope variation and relationship to detection of HCV RNA in blood. Hepatology 1999;29:1288-98.
• 34. Contreras AM, Ochoa-Jiménez RJ, Celis A, Méndez C, Olivares L, Rebolledo CE, et al. High antibody level: an accurate serologic marker of viremia in asymptomatic people with hepatitis C infection. Transfusion 2010;50:1335-43.
• 35. Sekongo YM, Kabore S, Dembele B, Yao KD, Bogui-Siransy L, Adjoumani JL, et al. Interest of confirmation tests in the diagnosis of viral hepatitis C to blood donors in Abidjan-Côte d'Ivoire. J Hematol Oncol Res 2020;3:32-7.
• 36. Lai KK, Jin M, Yuan S, Larson MF, Dominitz JA, Bankson DD. Improved reflexive testing algorithm for hepatitis C infection using signal-to-cutoff ratios of a hepatitis C virus antibody assay. Clin Chem 2011;57:1050-6.
• 37. Gong S, Schmotzer CL, Zhou L. Evaluation of quantitative real-time PCR as a Hepatitis C virus supplementary test after RIBA discontinuation. J Clin Lab Anal 2016;30:418-23.