Neisseria gonorrhoeae (GC) and Chlamydia trachomatis (CT) are the predominant causes of bacterial sexually transmitted diseases. While nucleic acid amplification testing (NAATs), primarily polymerase chain reaction (PCR), is regarded as the gold standard for identifying these two pathogens, it usually takes a prolonged turnaround time and requires sophisticated equipment with considerable expense. In this study, we developed novel loop-mediated isothermal amplification (LAMP) assays for rapid detection (< 30 min) of GC and CT in clinical urine and swab specimens. We analysed 208 clinical samples with three different pre-treatment techniques including heating inactivation, centrifugation, and DNA extraction. LAMP results were compared with clinical results from the FDA-approved BD ProbeTec ET assay. After heating inactivation, LAMP detected merely 41% and 65% of BD-identified GC- and CT-positive samples, respectively. Introducing centrifugation as an affordable and rapid pre-treatment step increased detection rates to 81% and 91% for GC and CT, respectively. DNA extraction further enhanced the detection rates to 96% and 95% for GC- and CT-LAMP, respectively. All these LAMP assays exhibited clinical specificity of ≥ 98%, underscoring the specificity of the chosen target genes (the porA pesudogene for GC and the ftsK gene for CT). Discrepant samples were verified by real-time PCR; results were consistent with our LAMP findings. The overall LAMP performance met the WHO criteria for sensitivity and specificity for GC/CT point-of-care testing.