Validation of Anti-Hiv 1/2 Confirmatory Immunoblot Assay and Development of Algorithm For Anti-Hiv Testing At A Specialized Healthcare Facility In Pakistan

Ali  Raza; Noor Fatima Shahid; Muhammad Numan; Anam Yousaf

doi:10.51253/pafmj.v76iSUPPL-5.13363

Authors

Ali Raza Pakistan Kidney and Liver Institute and Research Center Lahore, Pakistan
Noor Fatima Shahid Pakistan Kidney and Liver Institute and Research Center Lahore, Pakistan https://orcid.org/0000-0003-2402-0236
Muhammad Numan Pakistan Kidney and Liver Institute and Research Center Lahore, Pakistan
Anam Yousaf Pakistan Kidney and Liver Institute and Research Center Lahore, Pakistan

DOI:

https://doi.org/10.51253/pafmj.v76iSUPPL-5.13363

Keywords:

HIV testing, Anti-HIV antibody, Immunoblotting, Predictive Value of Tests Sensitivity and Specificity

Abstract

Objective: To validate the Anti-HIV 1/2 confirmatory, immunoblot assay and develop an optimized diagnostic algorithm and enhance Human Immunodeficiency Virus (HIV) detection by integrating serological and molecular methods.

Study Design: Cross-sectional analytical study.

Place and Duration of Study: Pakistan Kidney and Liver Institute and Research Center (PKLI&RC), Lahore, Pakistan, from Feb to Jul 2024.

Methodology: This study evaluated the diagnostic performance of the Anti-HIV 1/2 immunoblot assay and proposed a multi-step algorithm for HIV detection. Participants included hospitalized patients and blood donors. Initial screening were confirmed using Elecsys® HIV Combi PT assay through Electrochemiluminescence Immunoassay (ECLIA). Reactive samples were confirmed using the Bio-Rad Geenius™ HIV 1/2 confirmatory assay and Nucleic Acid Testing (NAT). Additional testing was performed using the Bioline™ HIV Immunochromatographic Test (ICT).

Results: Sixty participants were enrolled. Both immunoblot and ICT assays exhibited 100.00% sensitivity and specificity. NAT Polymerase Chain Reaction (PCR) demonstrated 100.00% sensitivity and 92.30% specificity, detecting acute infections missed by serological tests.

Conclusion: Integrating serological and molecular diagnostic improves HIV detection accuracy. The proposed algorithm, including ECLIA, immunoblot, and NAT, proved reliable. Modifying the ECLIA cutoff to ≥ 75.215 improved specificity from 57.70% to 100.00% while maintaining 100.00% sensitivity.

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