Background. Tuberculosis (TB) is an important public health concern in Nigeria. TB-associated stigma could lead to delayed diagnosis and care, treatment default, and multidrug resistance. Understanding of TB-associated stigma is therefore important for TB control. The study is aimed at determining and comparing the knowledge, attitude, and determinants of TB-associated stigma. Methodology. This was a comparative cross-sectional study among adults in urban and rural areas of Lagos State, Nigeria. Respondents were selected through a multistage sampling technique and interviewed using a semistructured questionnaire, which contained the Explanatory Model Interviewed Catalogue (EMIC) stigma scale. IBM SPSS Statistics Software package version 20 was used for analysis. Results. A total of 790 respondents were interviewed. High proportions of respondents in rural and urban areas were aware of TB (97.5% and 99.2%, respectively). Respondents in the urban areas had overall better knowledge of TB compared to the rural areas (59.4% vs. 23%;
p
<
0.001
), while respondents in the rural areas had a better attitude to TB (60.5% vs. 49.9%;
p
=
0.002
). The majority of respondents in rural and urban areas had TB-associated stigma (93% and 95.7%, respectively). The mean stigma score was higher in the urban compared to rural areas (
17.43
±
6.012
and
16.54
±
6.324
, respectively,
p
=
0.046
). Marital status and ethnicity were the predictors of TB-associated stigma in the rural communities (AOR-0.257; CI-0.086-0.761;
p
=
0.014
and AOR–3.09; CI-1.087-8.812;
p
=
0.034
, respectively), while average monthly income and age of respondents were the predictors of TB-associated stigma in urban areas (AOR–0.274; CI–0.009-0.807;
p
=
0.019
and AOR-0.212; CI–0.057-0.788;
p
=
0.021
, respectively). Conclusion. TB-associated stigma is prevalent in both rural and urban areas in this study. There is therefore a need to disseminate health appropriate information through the involvement of the community. Also, innovative stigma reduction activities are urgently needed.