Background: Lack of knowledge and understanding of standard precautions, and insufficient compliance with the established guidelines in nursing students will increase the risk of exposure to healthcare-associated infections (HAIs). This study aims to determine knowledge and compliance of standard precautions among the undergraduate nursing students and to identify the associated factors. Methods: A cross-sectional study was conducted among the undergraduate nursing students from a Malaysian public university in Pahang. A convenience sampling was used to select the participants. The participants were asked to complete an online survey consisting of questionnaire of knowledge and compliance of standard precautions. Results: A total of 178 undergraduate nursing students completed the questionnaires (response rate = 95%). Their mean age was 22.7 years old (SD = 1.1). The score of standard precautions knowledge was 90.9% (mean score = 16.4/18, SD = 1.5) and the score of compliance was 91.8% (mean score = 73.4/80, SD = 10.7). There were significant associations between age (p = 0.002), year of study (p = 0.032) and clinical posting experience (p = 0.010) with knowledge of standard precautions. Whilst only age (p = 0.011) was significantly associated with compliance of standard precautions. There was a significant positive association between knowledge and compliance of standard precautions (rs = 0.187, p = 0.012). Conclusion: Knowledge and compliance of standard precaution among the undergraduate nursing students were high and achieved the expected requirement for clinical posting. A continuous effort to improve adherence to standard precautions in nursing students is necessary to protect and prevent them from getting infected particularly during the COVID-19 pandemic.