Introduction Tuberculosis (TB) is the leading cause of death from a single infectious agent. Risk of TB disease is higher among people living with HIV. HIV prevalence among tuberculosis patients is in Africa is approximately 38%. Malawi registered a 44.6% TB/HIV coinfection in 2020. TPT prevents development of active TB disease. TPT uptake has been suboptimal Malawi. We conducted a study assessing the uptake of TPT and associated factors among PLWHIV in Kasungu district, Malawi. Methods A quantitative cross-sectional study among PLHIV from the 2021 cohort in Kasungu district. Target participants were from health facilities that provide TPT. An adapted questionnaire was used to collect socio-demographic factors including age, sex, education, residence, religion, marital status, economic status and assessed the knowledge of participants about TPT with their understanding of the use of TPT. The data was subjected to descriptive, bivariate and logistic regression analyses in SPSS version 25. Results Uptake of TPT was at 62%. Seventy five percent of the participants had good knowledge about TPT. Low uptake of TPT was associated with low education status, poor knowledge about TPT, being on ART for more than 24 months and receiving services at rural hospitals. Conclusion Low uptake of TPT needs to be addressed. There is for intensified health communication and promotion about TPT among the PLWHIV including diversifying the communication messages and channels to reach all literacy levels. Further studies are needed to ascertain other factors including the health care workers’ attitudes and perception about TPT.