This study aims to determine a suitable molecular identification technique for Fusarium oxysporum associated with tomato infections to enhance tomato production in developing countries. Solanum lycopersicum (tomato) is one of the most important vegetables worldwide and ranks as the sixth most popular vegetable, according to the Food and Agriculture Organization (FAO). However, its cultivation is significantly affected by pathogens, including F. oxysporum. The most suitable medium for cultivating F. oxysporum was found to be Potato Dextrose Agar (PDA). Cell disruption using bead beating in a homogenizer yielded optimal results. EF-1α primers were identified as the most appropriate to detect Fusarium isolates within species complexes, as corroborated by various researchers. Intergenic spacer restriction fragment length polymorphism (IGS-RFLP) analysis was widely used to trace the origin of F. oxysporum by analyzing genetic similarities among isolates from different sources. Other methods, such as random amplified polymorphic DNA (RAPD), restriction fragment length polymorphism (RFLP), allele-specific associated primer (ASAP), single nucleotide polymorphism (SNP), and variable number tandem repeat (VNTR) analysis have also been applied. To mitigate fungal infections in tomatoes, this study recommends selecting disease-resistant tomato varieties, maintaining ideal growing conditions, adhering to stringent sanitation practices, ensuring source water is free of potential fungal pathogens, sterilizing tomato seeds, and using organic fungicides as needed.