{Reference Type}: Journal Article {Title}: Hepatitis B Virus Prevalence among HIV-Uninfected People Living in Rural and Peri-Urban Areas in Botswana. {Author}: Anderson M;Mangogola T;Phinius BB;Mpebe G;Aimakhu CO;Choga WT;Phakedi B;Bhebhe LN;Ditshwanelo D;Baruti K;Mpofu-Dobo L;Othusitse L;Ratsoma T;Gaolathe T;Makhema J;Shapiro R;Lockman S;Moyo S;Gaseitsiwe S; {Journal}: Microorganisms {Volume}: 12 {Issue}: 6 {Year}: 2024 Jun 15 {Factor}: 4.926 {DOI}: 10.3390/microorganisms12061207 {Abstract}: (1) Background: we determined the prevalence of the hepatitis B virus (HBV) amongst people without human immunodeficiency virus (HIV) in rural and peri-urban areas in Botswana. (2) Methods: We screened for the hepatitis B surface antigen (HBsAg) from archived plasma samples of people without HIV (n = 2135) randomly selected from the Botswana Combination Prevention Program (BCPP) (2013-2018). We sequenced 415 bp of the surface region using BigDye sequencing chemistry. (3) Results: The median age of participants was 31 (IQR: 24-46) and 64% (1360/2135) were female. HBV prevalence was 4.0% (86/2135) [95% CI: 3.3-4.9]) and ranged between 0-9.2%. Older participants (>35 years) had increased odds of HBV positivity (OR: 1.94; 95% CI: [1.32-2.86]; p = 0.001). Thirteen samples were sequenced and seven (53.8%) were genotype A, three (23.1%) were genotype D and genotype E each. Clinically significant mutations were identified in the surface region, but no classic drug resistance mutations were identified. (4) Conclusions: We report an HBV prevalence of 4.0% (95% CI 3.3-4.9) among people without HIV in rural and peri-urban communities in Botswana with varying rates in different communities. A comprehensive national HBV program is required in Botswana to guide HBV prevention, testing and management.