Abstract: Background: Herpes simplex virus type 2 (HSV-2) and bacterial vaginosis are characterized by frequent reactivation and shedding of the virus and by the attendant risk of transmission to sexual partners. Aims and objective: to detection herpes simplex virus type two and some bacterial vaginosis from women and inductions of these microbes with them due to chronic infection. Patients and methods: This Cross Sectional study was carried out for a period of (1) year from July (2019) to July (2020). This study included 150 women were suffering from bacterial vaginosis the sample was high vaginal swabs from married female were admitted to the out-patient clinics of Gynecology and Babylon Hospital for Maternity and Pediatrics. End cervical swab or secretion and blood from each case were collected following standard procedure for microscopic examination and isolation of bacteria. All the isolates were screened and identified via the VITEK-2 System (BioMerieux). This is a phenotypic type of identification, which depends on biochemical reactions to identify the isolates. Herpes simplex virus was detected by immunofluorescence assay and qRCR. Results: In this study, 150 vaginal swab positive growth after culturing (aerobic and anaerobic) on different media and detected on GN-ID, GP-ID with VITEK-2 Compact were identified many types of isolated bacteria were found, these isolates were included 35(23.3%) Streptococcus viridians, 30(20%) Gardnerella vaginitis, 25(16.7%) Lactobacillus, 25(16.7%) Listeria monocytogens, 10(6.7%) Bacteroides, 10(6.7%) Prevottella, 8(5.3%) Fusobacteriam and 7(4.6%) Enterococcus faecalis. In this study revealed that the Immunofluorescence assay detect 55/150 (36.6%) of cases positive for Herpes simplex Virus type 2. Herpes simplex Virus type 2 was isolated from blood samples by qPCR technique. It was found that, from (150) samples of women vaginosis, 55(36.6%) isolated were related of Herpes simplex Virus type 2 at 272 bp. Conclusions: Herpes simplex virus type 2 (HSV-2) and bacterial vaginosis are two of the most common important infections of the female genital tract, both are associated with an increased risk for acquiring HIV infection, and they are associated with each other.