Demodex sp. is a skin ectoparasite known as follicle mite. The objectives of this study were to determine the prevalence of Demodex sp. infestation among Malaysian adults with age ranges from 20 to 29 years old in Lembah Klang and to determine the relationship between the infestation status and gender and acne problem on face. This is the first study that has been conducted in Malaysia to determine the prevalence of Demodex sp. infestation. A total of 350 people participated in this study. Samples were selected by stratified random sampling. Skin scraping was carried out on forehead, nose, cheeks and chin. All specimens were mounted in Hoyer’s medium and studied under light microscope. Questionnaires were distributed to the subjects to identify acne problem on their skin conditions. The overall prevalence of Demodex sp. infestation was 18.9% with higher prevalence in males (25.7%) compared to females (12.0%). There was no significant difference between acne problem and Demodex sp. infestation. In conclusion, infestation of Demodex sp. was detected in a fifth of the subjects, with men had more problems.