COMPARATIVE ANALYSIS OF SEROLOGICAL AND MOLECULAR TRANSFUSION TRANSMISSIBLE INFECTIONS SCREENING

Marija Tashkovska; Tatjana Makarovska Bojadjieva; Emilija Velkova; Violeta Dejanova Ilijevska; Elena Petkovic; Elena Ristovska; Bojan Todorovski; Ivana Babalj Veljanoski; Aleksandra Stojchevska

Marija Tashkovska City General Hospital 8th September, Skopje, North Macedonia
Tatjana Makarovska Bojadjieva Institute of Transfusion Medicine, Faculty of Medicine, Ss. Cyril and Methodius University in Skopje, North Macedonia
Emilija Velkova Institute of Transfusion Medicine, Faculty of Medicine, Ss. Cyril and Methodius University in Skopje, North Macedonia
Violeta Dejanova Ilijevska Institute of Transfusion Medicine, Faculty of Medicine, Ss. Cyril and Methodius University in Skopje, North Macedonia
Elena Petkovic Institute of Transfusion Medicine, Faculty of Medicine, Ss. Cyril and Methodius University in Skopje, North Macedonia
Elena Ristovska Institute of Transfusion Medicine, Faculty of Medicine, Ss. Cyril and Methodius University in Skopje, North Macedonia
Bojan Todorovski Institute of Transfusion Medicine, Faculty of Medicine, Ss. Cyril and Methodius University in Skopje, North Macedonia
Ivana Babalj Veljanoski City General Hospital 8th September, Skopje, North Macedonia
Aleksandra Stojchevska City General Hospital 8th September, Skopje, North Macedonia

Abstract

Introduction. Transfusion Transmissible Infections (TTIs) remain a major concern in ensuring the blood safety. Nucleic acid testing (NAT) improves detection by shortening the window period (WP) and identifying occult infections. This study aims to determine the significance of NAT through a comparative analysis of the chemiluminescent serological technique (CMIA) and the molecular technique (NAT). Methods. This retrospective study analyzed TTI testing data from 167979 blood donations (93,390 donors) between 2022 and 2024. Screening included HBV DNA, HCV RNA, and HIV RNA (Procleix UltrioPlex E, Panther System) and serological markers (HBsAg, anti-HCV, anti-HIV/p24) using the Architect 2000 platform. Repeatedly seroreactive samples underwent confirmatory testing (HBsAg neutralization, HCV/HIV immunoblot), while NAT-reactive samples were individually tested using the Procleix Ultrio Elite discriminatory assay. Results. Serological testing identified 237 (0.25%) confirmed TTI cases, while NAT detected 245 (0.26%), including 22 (0.023%) NAT-only positive cases. Among these, 21 were HBV DNA-positive/HBsAg-negative. Follow-up samples from 11 donors showed seroconversion in 7 (33.3%), while 4 (19.0%) were identified as potential occult HBV infections (OBI). One HIV NAT-only case was detected. The NAT yield was 1 per 4,245 donations, decreasing over time. Residual risk per 100,000 donations declined from 10.55 to 4.59 for HBV, while remaining low for HCV and HIV (0.12 in 2024). Conclusion. NAT enhances blood safety by detecting early and occult infections missed by serology. The declining NAT yield and residual risk emphasize improved screening efficiency. Integrating NAT with serological methods is crucial for reducing transfusion-transmitted infections.

Keywords: NAT, blood safety, transfusion-transmissible infections, residual risk

Author Biography

Emilija Velkova, Institute of Transfusion Medicine, Faculty of Medicine, Ss. Cyril and Methodius University in Skopje, North Macedonia

References

1. Busch MP, Bloch EM, Kleinman S. Prevention of transfusion-transmitted infections. Blood. 2019 Apr 25;133(17):1854-1864. doi: 10.1182/blood-2018-11-833996. Epub 2019 Feb 26. PMID: 30808637.
2. Bihl, F., Castelli, D., Marincola, F. et al. Transfusion-transmitted infections. J Transl Med 5, 25 (2007). https://doi.org/10.1186/1479-5876-5-25
3. World Health Organization (WHO). Screening Donated Blood for Transfusion-Transmissible Infections: Recommendations. Geneva: WHO; 2009. Available from: https://www.ncbi.nlm.nih.gov/books/NBK142989/.
4. Kim HJ, Ko DH. Transfusion-transmitted infections. Blood Res. 2024. 12;59(1):14. doi: 10.1007/s44313-024-00014-w. PMID: 38607595; PMCID: PMC11014835.
5. Hans R, Marwaha N. Nucleic acid testing-benefits and constraints. Asian J Transfus Sci. 2014 Jan;8(1):2-3. doi: 10.4103/0973-6247.126679. PMID: 24678164; PMCID: PMC3943139.
6. Laperche S. Blood safety and nucleic acid testing in Europe. Euro Surveill. 2005 Feb;10(2):3-4. PMID: 15735316.
7. Hans R, Marwaha N. Nucleic acid testing-benefits and constraints. Asian J Transfus Sci. 2014 Jan;8(1):2-3. doi: 10.4103/0973-6247.126679. PMID: 24678164; PMCID: PMC3943139.
8. Schmitz JE,,Stratton CW,Persing DH, Tang Y.2022.Forty Years of Molecular Diagnostics for Infectious Diseases. J Clin Microbiol60:e02446-21.https://doi.org/10.1128/jcm.02446-21
9. World Health Organization. WHO guidelines on estimation of residual risk of HIV, HBV or HCV infections via cellular blood components and plasma [Internet]. Geneva: WHO; [cited 2023 Apr 1]. Available from: https://cdn.who.int/media/docs/default-source/biologicals/blood-products/document-migration/resriskgl_who_trs_1004_web_annex_4.pdf?sfvrsn=55dd09d3_3.
10. Liang TJ. Hepatitis B: the virus and disease. Hepatology. 2009 May;49(5 Suppl):S13-21. doi: 10.1002/hep.22881. PMID: 19399811; PMCID: PMC2809016.
11. .WHO Hepatitis B. [(accessed on januar 2025)]. Available online: https://www.who.int/news-room/fact-sheets/detail/hepatitis-b.
12. Asandem DA, Segbefia SP, Kusi KA, Bonney JHK. Hepatitis B Virus Infection: A Mini Review. Viruses. 2024 May 3;16(5):724. doi: 10.3390/v16050724. PMID: 38793606; PMCID: PMC11125943.
13. Raimondo G, Pollicino T, Cacciola I, Squadrito G. Occult hepatitis B virus infection. J Hepatol. 2007 Jan;46(1):160-70. doi: 10.1016/j.jhep.2006.10.007. Epub 2006 Nov 7. PMID: 17112622.
14. Candotti D, Boizeau L, Laperche S. Occult hepatitis B infection and transfusion-transmission risk. Transfus Clin Biol. 2017 Sep;24(3):189-195. doi: 10.1016/j.tracli.2017.06.014. Epub 2017 Jun 30. PMID: 28673499.
15. Gerlich WH, Bremer C, Saniewski M, Schüttler CG, Wend UC, Willems WR, Glebe D. Occult hepatitis B virus infection: detection and significance. Dig Dis. 2010;28(1):116-25. doi: 10.1159/000282074. Epub 2010 May 7. PMID: 20460899.
16. Kiely P, Margaritis AR, Seed CR, Yang H; Australian Red Cross Blood Service NAT Study Group. Hepatitis B virus nucleic acid amplification testing of Australian blood donors highlights the complexity of confirming occult hepatitis B virus infection. Transfusion. 2014 Aug;54(8):2084-91. doi: 10.1111/trf.12556. Epub 2014 Mar 20. PMID: 24650170.
17. Basit H, Tyagi I, Koirala J. Hepatitis C. 2022 Nov 26. In: StatPearls [Internet]. Treasure Island (FL): StatPearls Publishing; 2023 Jan–. PMID: 28613647.
18. Messina JP, Humphreys I, Flaxman A, Brown A, Cooke GS, Pybus OG, Barnes E. Global distribution and prevalence of hepatitis C virus genotypes. Hepatology. 2015 Jan;61(1):77-87. doi: 10.1002/hep.27259. Epub 2014 Jul 28. PMID: 25069599; PMCID: PMC4303918.
19. World Health Organization (WHO). Hepatitis C. [ Apr; 2024 ]. 2024. https://www.who.int/news-room/fact-sheets/detail/hepatitis-c https://www.who.int/news-room/fact-sheets/detail/hepatitis-c
20. Manns MP, Maasoumy B. Breakthroughs in hepatitis C research: from discovery to cure. Nat Rev Gastroenterol Hepatol. 2022 Aug;19(8):533-550. doi: 10.1038/s41575-022-00608-8. Epub 2022 May 20. PMID: 35595834; PMCID: PMC9122245.
21. Kumar A, Rajput MK, Paliwal D, Yadav A, Chhabra R, Singh S. Genotyping & diagnostic methods for hepatitis C virus: A need of low-resource countries. Indian J Med Res. 2018 May;147(5):445-455. doi: 10.4103/ijmr.IJMR_1850_16. PMID: 30082568; PMCID: PMC6094507.
22. World Health Organization. Combating hepatitis B and C to reach elimination by 2030 [Internet]. Geneva: WHO; 2016 [cited 2023 Apr 1]. Available from: https://apps.who.int/iris/handle/10665/206453.
23. World Health Organization. HIV/AIDS fact sheet [Internet]. Geneva: WHO; [cited 2023 Apr 1]. Available from: https://www.who.int/news-room/fact-sheets/detail/hiv-aids.
24. Eberle J, Gürtler L. HIV types, groups, subtypes and recombinant forms: errors in replication, selection pressure and quasispecies. Intervirology. 2012;55(2):79-83. doi: 10.1159/000331993. Epub 2012 Jan 24. PMID: 22286874.
25. Fearon M. The laboratory diagnosis of HIV infections. Can J Infect Dis Med Microbiol. 2005 Jan;16(1):26-30. doi: 10.1155/2005/515063. PMID: 18159524; PMCID: PMC2095005.
26. Levy JA. Pathogenesis of human immunodeficiency virus infection. Microbiol Rev. 1993 Mar;57(1):183-289. doi: 10.1128/mr.57.1.183-289.1993. PMID: 8464405; PMCID: PMC372905.
27. World Health Organization. WHO Guidelines on Estimation of Residual Risk of HIV, HBV or HCV Infections via Cellular Blood Components and Plasma. Expert Committee on Biological Standardization, Geneva, 17–21 October 2016. Available from:www.who.int/biologicals/expert_committee/Residual_Risk_Guidelines_final.pdf [Accessed: February 9, 2025].