Identification of Lactobacillus Species from Vaginal Secretions of Healthy and Women with Vaginitis by Molecular Methods

Background: The Lactobacillus species bear a high responsibility toward vaginal microbiota and, accordingly, play a very important role in the maintenance of vaginal health. In healthy women, Lactobacillus bacteria keep the vagina in an acid environment and helps in maintaining the microbiota balance. Objective: To identify and compare Lactobacillus species in vaginal secretions of healthy women and those with vaginitis, by using molecular techniques. Methodology: The study was conducted in College of Medicine laboratories- University of Anbar and Private Specialized laboratories from November 2023 until October-2024. 234 specimens were collected from women with expected vaginitis, their ages ranged from 20-50 years they were selected from Al-Ramadi Teaching Hospital for maternity and pediatrics in Ramadi, Western Iraq. Genomic DNA extraction from vaginal secretions was performed using Presto™ Mini gDNA Bacteria Kit. The Nano drop instrument was used to calculate DNA purity and concentration. Multiplex Polymerase chain reaction (PCR) was carried out using Multiplex-PCR primer design for ten of Lactobacillus species. All Multiplex-PCR reactions were done in Applied Bio-system 2720 thermo cyclers, and amplification of target gene. Results: The results of sampling show that 148/234 of specimens were identified a vaginitis positive while 86/234 were negative. The molecular screening findings by Multiplex-PCR indicated that five species were identified as L. jensenii, L. acidophilusL. crispatus,L. iners, L. gasseri as common species in the vaginal microbiome. Conclusion: The analysis results show that the more frequent species in women case was L. crispatus comparing to the control group with p-value was 0.001, they were present in 26 samples of patients versus 11 samples of controls. The consistent presence of Lactobacillus iners across different types of vaginitis is a notable finding