Designing a PCR-Based Genotyping Technique for the LEP A19G Polymorphism: Method Development and Optimization

Abstract

The LEP gene is a protein-encoding gene for the satiety hormone called leptin, which stimulates appetite, impeding food intake and anorexigenic peptide upregulation. This research aims to develop a genotyping method to detect the LEP gene variant A19G, known for its association with appetite and eating preference. Convenience random sampling was performed by disseminating the subject's recruitment on social media among the students of Faculty of Health Sciences, Universiti Teknologi MARA. A total of 51 undergraduates (mean age, 23.20 (Standard Deviation, SD 1.56) years old; 41 females, 10 males) from the faculty were recruited. Questionnaires have been provided to assess the subjects' meal preferences and food intake frequency. A19G genotyping was performed using polymerase chain reaction (PCR) and subsequent agarose gel electrophoresis (AGE). The analysis confirmed the successful development of the A19G genotyping method. The variant allele frequency (VAF) for A19G was 0.13. This approach provides a reliable genotyping tool for LEP A19G in future investigations despite data suggesting that A19G may not be a marker for appetite differences in this subject group. Further research with a larger, more diverse population is warranted to confirm these results and explore the influence of other LEP gene variants on appetite and eating preferences.