Вагинален микробиом кај женска популација во репродуктивен период

Abstract

The molecular analysis of the vaginal microbiome (VM) is one of the most interesting current issues in the field of reproductive health and perinatal medicine, as a basis for developing new strategies for prediction, prevention and treatment of genital infections. OBJECTIVES: Genetical determination of the Lactobacillus Species in vaginal discharge and the most common causes of bacterial vaginosis in our population, i.e. molecular analysis of the vaginal microbiome in terms of the types of Lactobacillus, among women with normal findings, intermediate flora and bacterial vaginosis, proved with standard microbiological and clinical parameters. To compare the clinical parameters and standard microbiological analysis for bacterial vaginosis with molecular tests and to determine the possible causes. To make an assessment for development of a simple method for analysis of the vaginal microbiome. MATERIAL AND METHODS: Prospective study of 110 reproductive-age women who referred for gynecological examination at CH Acibadem Sistina. The vaginal examination included measurement of the vaginal pH and taking two samples, with sterile swab, from the vaginal fornices, for bacteriological and molecular analysis. The first sample was used for the Nugent score analysis and divided the women in three groups: normal finding (0-3); intermediate flora (4-6) and bacterial vaginosis (7-10). The sample for molecular analysis was immediately frozen at -20 ° C and thus transported to the Laboratory at the Academy of Sciences and Arts of Republic of North Macedonia. 93 samples were selected randomly or molecular analysis of Lactobacillus spp, Gardnerella vaginalis, Mycoplasma spp and Ureaplasma spp. Identification was done by amplification of 16S rRNK genes. The complete 16S rRNK gene (1.5 kb) was amplified by PCR using primers and sequenced. Sequences were compared for identification of isolates. Based on the differences, a genealogical tree- dendrogram was created. Identification is considered to be confirmed at 99-100% homology to the type strain sequence. RESULTS: Colonization with Lactobacillus significantly associates with normal vaginal flora compared with bacterial vaginosis. 22 Species of Lactobacillus have been detected, most commonly Lactobacillus iners, Lactobacillus crispatus and Lactobacillus casei. Colonization with Lactobacillus crispatus and Lactobacillus casei significantly associates with normal vaginal flora and reduces the chance of registration of bacterial vaginosis. The presence of Lactobacilli has been verified in all women with normal findings. There was a statistically significant difference in the percentage of Lactobacilli in the group with normal flora compared to bacterial vaginosis, as well as in the group with intermediate finding versus bacterial vaginosis. Molecular analysis confirmed the presence of causative agents of bacterial vaginosis. An overview of the types of lactobacilli was obtained, and a dendrogram was created. There was a significant correlation of clinical parameters and classical microbiological findings for bacterial vaginosis with molecular markers.