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Background: Sepsis is a major cause of maternal death worldwide and caused by a variety of bacterial pathogens. In many developing countries including Ethiopia, the treatment of puerperal sepsis is based on empirical/syndromic that may promote antimicrobial resistance. Therefore the aim of this study was to investigate bacterial pathogens, their antimicrobial susceptibility patterns, and associated factors among women with suspected puerperal sepsis. Methods: A cross-sectional study was conducted among puerperal sepsis suspected women attending Asella Referral and Teaching Hospital, from September 2020 to August 2021. A total of 174 study participants were enrolled. Sociodemographic and obstetric data of the participants were collected using a pretested structured questionnaire and checklist respectively. About 20 ml blood sample was collected from all study participants into BacT/ALERT® 3D blood culture bottles and incubated into BacT/ALERT® 3D automated blood culture system. Endocervical swab was also collected into Aime's transport media. Bacterial isolation and identification was done using standard bacteriological methods. Disc diffusion method was used to determine the antimicrobial susceptibility profiles of bacterial isolates. Data were entered into EpiData version 4.6 and transferred to SPSS version 25.0 for analysis. Results: The overall positivity rate of bacterial isolates among puerperal sepsis suspected women was 48.9%. Out of these 87.1% of the isolates were Gram negative bacteria. The most common isolates were E. coli (54.1%) followed by Klebsiella spp. (23.5%) and S. aureus (10.6%). E. coli showed a higher resistance rate to Piperacillin (87%). Klebsiella spp. showed a higher resistance rate to Aztreonam (65%) and Ceftriaxone (65%). S. aureus showed a higher resistance rate to Trimethoprim-sulfamethoxazole (66.6%). In this study, 81.2% of the isolates were multi-drug resistant bacterial pathogens. Multivariate regression analysis showed no statistically significant association between sociodemographic, obstetrics factors, and having bacteria. Conclusion: In this study the overall positivity rate in this study was 48.9%. E. coli, Klebsiella species, and S. aureus were the most common isolated bacteria. High numbers of multidrug-resistant bacterial isolates were identified. Our finding emphasizes the need for strengthening microbiology services for better management of patients.