Bacterial vaginitis is one of the common conditions in the reproductive age of women characterized by inflammation in the vaginal mucosa. Among the various etiological agents that influence vaginitis, one of the most common etiological agents is bacteria that belongs to Enterobacteriaceae family members, including Escherichia coli and Klebsiella pneumoniae, which have been found as the primary common pathogen of aerobic vaginitis. This study aims to determine the genetic relatedness of the Enterobacterial Repetitive Intergenic Consensus (ERIC-PCR) technique isolated from pregnant and nonpregnant patients diagnosed with bacterial vaginosis.The patient's vaginal swabs were collected using a sterile vaginal swab and screened for Gram-negative bacteria, and then the genus of those bacteria was identified using gold-standard microbiology techniques such as culturomics. The disc diffusion method was used to determine the antibiotic susceptibility of the bacteria to extended-spectrum beta-lactamase (ESBL). The organism's susceptibility was tested against eleven antimicrobial agents. A single-plex PCR was carried out for the following genes: Temoneira (TEM), Sulfhydryl reagent variable (SHV), and Cefotaxime-hydrolyzing β-lactamase (CTXM). After identifying ESBL resistance using the endpoint PCR, the genetic relatedness between each strain was determined using ERIC-PCR. Then, the gel was analyzed using the Gel-J software to create the phylogenetic tree dendrogram to find the genetic variations.Antibiotic susceptibility testing and molecular detection of antibiotic resistance genes demonstrated that antibiotic resistance is more prevalent in E. coli and K. pneumonia, which was shown to be the primary causative agent involved in bacterial vaginosis towards fluoroquinolone resistance. Over fifty percent of the isolates exhibited a multidrug resistance trait.This study's findings demonstrate an increase in multi-resistant strains of K. pneumoniae and E. coli prevalent in pregnant and nonpregnant women after examination. The results of the ERIC PCR analysis showed a significant genetic diversity between the strains of K. pneumoniae and E. coli, indicating the polyclonal distribution of these isolates in both pregnant and nonpregnant women presented with vaginal infections.