Abstract

MULTIPLEX PCR TECHNIQUE TO IDENTIFY OF SUDANESE FEMALES VAGINAL MICROBIOTA

Deena A. Mohamed*, Einas A Osman, Sababil S. Ali, Asim B. Abdel-Mageed and Hassan A. Aziz

ABSTRACT

Background: Microbiota of healthy women changes from birth to menopause. During their reproductive years, female’s normal flora is dominated by Lactobacillus species, females whom are identified with vaginal infections are characterized by lacking Lactobacillus species and the community is composed of facultative and strictly anaerobic bacteria. Recently, a new level of detailed description of the vaginal microbiome has been attained using quantitative, real-time, PCR targeting key bacterial genomes. In this study we have used 16S RNA sequencing to identify the Sudanese flora within females in their reproductive age. Objectives: Molecular characterization of the vaginal microbiome among Sudanese females within the reproductive age and the micro-flora associated with bacterial vaginosis (BV) in Khartoum State, Sudan using molecular diagnosis (conventional PCR technique) of vaginal samples. Materials and Methods: This is a descriptive cross sectional and molecular laboratory based study; was carried out in Khartoum state-Sudan selected two major reproductive centres. Vaginal swab samples were collected from participants who visited reproductive centres in period between March 2016 to March 2017. The DNA was then extracted from the swab samples and 16S RNA sequencing was performed. Results: Multiplex PCR screened for 100 vaginal swabs samples. Lactobacillus species was found to be the most dominant organism in all specimens except of samples with bacterial vaginosis (B.V) which was found to be dominated with the genus Gardnrella. Contraceptive and skin smoking (Dukhan) were found to have a significant impact on the presence of vaginal flora with P-value of 0.000 and 0.003 respectively. Conclusions and Recommendations: Sudanese female’s flora is mostly dominated by Lactobacillus species among normal and asymptomatic females and Gardnrellla sp. Among females with B.V. more studies and metagenomics sequencing is recommended for better identification.

Keywords: microbiota, PCR, Lacbacillus, Bacterial Vaginosis.