Molecular Identification and Genotyping of Atopobium Vaginae, 16S rRNA Gene from Bacterial Vaginosis Miscarriage Women in Al-Hillah City

Abstract

This study was aimed to determine the Atopobium vaginae associated bacterial vaginosis (BV) in vaginosis women and women with miscarriage. Also other aim, the deoxyribonucleic acid (DNA) sequencing was performed for phylogenetic tree analysis of 16S rRNA gene in local A. vaginae isolates in comparison with NCBI-Genbank global A. vaginae isolates and finally submission of the present isolates in NCBI-Genbank database. One hundred fifty high vaginal swabs were collected from women with vaginosis(75 samples were taken from married vaginosis women without miscarriage and 75 samples from vaginosis women with miscarriage) from Babylon city hospital and private clinics. The age of patient 15 to 45 years. The sample was collected by disposable swabs, genomic DNA was extracted from these swabs. 16S rRNA gene detection by polymerase chain reaction technique . A. vaginae was isolated on Columbia blood agar supplemented with antibiotic for the first time in Iraq, the study confirmed that 9 (12.00%) and 5 (6.66%) of A. vaginae out of 150 swabs isolated from miscarriage and non-miscarriage vaginosis women respectively. According to the detection of the 16S rRNA gene, the study revealed that 69 (92.00%) and 47(62.66%) of A. vaginae out of 150 swabs obtained from miscarriage and non-miscarriage vaginosis women respectively. Basic Local Alignment Search Tool (BLAST) analysis showed that the 16S rRNA gene shared more than 98–99% sequence compatibility with the sequences of A. vaginae. Furthermore, the phylogenetic tree analysis of the 16S rRNA gene indicated that local A. vaginae (NO.1 and NO. 2 ) isolates shared higher homology with other A. vaginae isolates available in the GenBank. The homology of the nucleotides was between (99.17 and 98.75%) respectively.