Chlamydia trachomatis, a major cause of sexually transmitted infection, poses a range of symptoms including genital discharge, pain during urination, and abdominal pains in women, and can lead to serious health complications such as pelvic inflammatory diseases, infertility, and ectopic pregnancy if left untreated. The need for rapid and accurate detection is imperative so prompt treatment and control of the disease can be achieved. This study conducted an immunoinformatic analysis of proteins of Chlamydia trachomatis (incA, hctA, ompA, omcB, rpoB, and HSP60) for the development of a lateral flow assay-based diagnostic test. Detailed in silico evaluation of selected proteins from publicly available genomic databases was conducted to evaluate their suitability as targets for lateral flow assay-based detection. The series of tests included antigenicity, toxicity, solubility, physicochemical characteristics and molecular docking of the derived constructs, and protein sequence. Chimeric construct was derived from the prediction of linear B cell epitopes, helper T cell major histocompatibility complex II binding epitopes, and IL4 and IL10 inducers using bioinformatic tools at standard thresholds. With a Ramachandra’s score of 95.4% and Z-score of -5.1, results indicate that the construct efficacy is high in potential to provide extreme specificity and sensitivity for the detection of Chlamydia trachomatis in clinical samples as compared to traditional culture-based methods using nucleic acid amplification, hereby providing a quicker and more accurate diagnostic tool for Chlamydia trachomatis infection. Findings offer valuable data for the development of a rapid and reliable diagnostic point-of-care test kit for Chlamydia trachomatis that allows for drastic reduction in clinical wait time and treatment.