Integrated laboratory protocol for the diagnosis of Sexually Transmitted Infections (STIs): Standardized pre-analytical procedures, rapid screening, hemagglutination, and ELISA methods for use in resource-limited settings

Abstract

This laboratory protocol describes a fully standardized and integrated diagnostic workflow for the detection of major sexually transmitted infections (STIs), including HIV-1/2, Treponema pallidum, hepatitis B virus (HBV), hepatitis C virus (HCV), Chlamydia trachomatis , and HSV-2. The workflow combines rapid immunochromatographic tests, non-treponemal and treponemal assays, hemagglutination testing, and ELISA-based antibody and antigen detection into a coherent algorithm. The protocol is aligned with ISO 15189 and Clinical and Laboratory Standards Institute (CLSI) recommendations and is specifically designed to be feasible in resource-limited laboratories. It includes detailed pre-analytical requirements for blood collection, transport, and storage; standardized step-by-step procedures for each assay; internal and external quality assurance components; troubleshooting guidance; and recommendations for data management and sample traceability. This integrated approach aims to optimize diagnostic yield, ensure reproducibility, and support large-scale epidemiological studies or routine diagnostic activities in low-income settings where access to molecular testing is limited.