A Comparative Assessment of Immunochromatography, Elisa and PCR for the Detection of Hepatitis B Surface Antigen in Human Plasma

Awadh Sahu, Naresh Kumar Dewangan, Sumit Gupta

Abstract: Hepatitis B virus (HBV) is a common blood borne infectious agents causing high morbidity and mortality that constitute the major global health challenge These viruses are mainly spread through the translation of contaminated blood and blood products. HBV is a DNA virus belonging to the Hepadnaviridae family The primary laboratory screening test for the early detection of HBV infection is the immunoassay for hepatitis B surface antigen (HBsAg). Various advanced analytical techniques are currently employed for HBV diagnosis, including Immunochromatographic Test (ICT), Enzyme-Linked Immunosorbent Assay (ELISA), Chemiluminescent Microparticle Immunoassay (CMIA), and Polymerase Chain Reaction (PCR). Findings from various studies indicate that rapid testing methods may not always ensure the accuracy of results, thereby increasing the risk of bloodborne infections during transfusion. In the early stages of infection or during the recovery phase, viral titers are often low, leading to reduced signal optical density (Low SOD) in test outcomes. These low-positive cases may go undetected by rapid screening methods such as ICT but can be identified through more sensitive assays like ELISA. and PCR However, it is essential to emphasize that standardization of diagnostic methods is critical before confirming an infection. A key consideration in the use of screening tests is that they must demonstrate high diagnostic performance across all stages of the disease to ensure reliable detection. In the present study, a comparison was done between ELISA, PCR, CLIA and ICTs for the screening of HBsAg. This study aims at comparing the analytical measures between ELISA, PCR, CLIA and rapid cards for better diagnosis of HBsAg in forthcoming years. From this study results, it was found that for HBsAg screening, ELISA, PCR, CLIA was more sensitive than the rapid card tests and equally specific to the rapid card tests. In this study, the sensitivity of ICT was 83.4%,specificity 100%, PPV 99.4%, NPV 100 % and the sensitivity, specificity, PPV and NPV ELISA was 100%.(Prabha, P. et al.2022), the sensitivity of ICT was 97 %,specificity 100%, PPV 90.4 %, NPV 100 % and the sensitivity, specificity, PPV and NPV ELISA was 100%.(Rahaman, S. et al. 2023), the sensitivity of ICT was 80%,specificity 100%, PPV 97.8.%, NPV 100 % (Chameera, E.W.S. et al.2013), the sensitivity of ICT was 97.1%,specificity 100%, PPV 98.5.%, NPV 100 % and the sensitivity, specificity, PPV and NPV PCR was 100% (Hasan, K. N.et al. 2017), the sensitivity of ELISA was 78%,specificity 76 %, PPV 75.%, NPV 80% and the sensitivity, specificity, PPV and NPV PCR was 100% (Navvabi, N. et al. 2021) , the sensitivity of ELISA was 100%,specificity 100%, PPV 100%, NPV 100% and the sensitivity, specificity, PPV and NPV TRUE NAAT PCR was 100% (Samom, P., et al .2023) ,the sensitivity of ELISA was 92%,specificity 100%, PPV 100.%, NPV 97% and the sensitivity, specificity, PPV and NPV PCR was 100% (Aftab, H. T.,et al. 2023) ,the sensitivity of ICT was 92%,specificity 97%, PPV 93%, NPV 96 % and the ELISA was sensitivity95%, specificity99.82%, PPV95% and NPV 99% (Ali, I., et al. 2024) ,the sensitivity of ICT was 98%,specificity 100%, PPV 100.%, (Hayder, I., et al. 2012), the sensitivity of ICT was 82%, specificity 100%, PPV 91%, NPV 100 % and the sensitivity, specificity, PPV and NPV ELISA was 100% (Asaduzzaman, M., et al. 2012), the sensitivity of ICT was 100%, specificity 100%, PPV 100%, NPV 100 % and the sensitivity, specificity, PPV and NPV ELISA was 100%. and sensitivity, specificity, PPV and NPV PCR was 100%. (Habibi, A., et al. 2016) ,the sensitivity of ICT was 99.7%,specificity 100%, PPV98% (Al-Matary, A. M., et al. 2016) , the sensitivity of ICT was92.98%,specificity 100%, PPV 99.4%, NPV 94.73 % and the sensitivity CLIA was 100%, specificity100%, PPV100% and NPV 97% and the sensitivity, specificity, PPV and NPV PCR was 100% (RENGARAJ, R., et al. 2024) ,the sensitivity of ICT was 99.27%,specificity 85.52%, PPV 95.58%, NPV 97.38 % and the ELISA was sensitivity 98.79%, specificity89.04%, PPV92.85% and NPV98.08% and sensitivity, specificity, PPV and NPV PCR was 100%. (Irfan, S., et al. 2018), the sensitivity of CLIA was 96.77%, specificity 96.07% PPV 93.75%, NPV 98% and the sensitivity, specificity, PPV and NPV ELISA was 100%., (Majumder, P., et al. 2017). This study does not have 100 % in the sensitivity and specification of the rapid test, but still some good brand rapid kits also have 100 % sensitive and specification, while the rapid test can also be done in rural areas and small labs, but after getting positive in the Rapid Test, it should be confermed with other mathods.

Keywords: HBsAg diagnosis, ELISA vs ICT, hepatitis B screening, diagnostic test sensitivity, blood transfusion safety