Comparison of Bis NEG-D and API 20E for the Identification of Gram-negative Bacilli in the Laboratory of the University Hospital of Befelatanana Antananarivo Madagascar

In medical analysis laboratories, techniques for identifying bacteria are currently becoming more and more numerous. The objective of this study is to compare the 2 bacterial identification systems Bis NEG-D and Api 20E for the identification of gram-negative bacilli. This is a qualitative evaluation of the Bis NEG-D compared to the gold standard Api 20E. During the study period, 32 Gram-negative bacilli isolates were identified simultaneously using Api 20E and Bis NEG-D. The samples are represented by 12 (37.5%) blood samples for blood culture, 12 (37.5%) urine samples for cytobacteriological examination of the urine, 06 (18.8%) pus samples for bacteriological examination of pus, a sample of the cerebrospinal fluid (3.1%) and a vaginal sample (3.1%).The bacteria identified were represented by Enterobacter spp, Escherichia col,i Klebsiella pneumonia, Shigella spp, Salmonella typhi, Acinetobacter baumannii, Pseudomonas spp, Proteus mirabilis, Raoultella ornithinolytica and Bukholderia cepacia. This study showed a concordance of 90.6% (29/32) and a discordance of 9.4% (3/32) between the results of Api 20E and Bis NEG-D. Concerning the probability scores, they vary between 95% to 100% for Api 20E and between 79.3% to 100% for Bis NEG-D. This study also compared the pros and cons of using Api 20 E and Bis NEG-D. The Bis-NEG-D is valid and can be used by medical analysis laboratories like the Api 20E, especially if these laboratories do not need to perform a lot of bacterial identification tests.

Identification of female genital tract infectious agents in patients treated for cytopathological diagnosis

Vaginal candidiasis, fungal infection caused by species of Candida spp. that affects women of all ages, cultures, habits, social and economic conditions. The present study had as its main objective to determine the identification of Candida spp. isolated in a vaginal sample, collected together with the collection for the cytopathological exam, in women treated in a university extension project, aiming at allowing a suitable therapeutics afterwards. The extension project began in January 2014 to June 2019 at the UFRGS. The patients who sought the Laboratory of Clinical and Toxicological Analysis (LACT) of the Faculty of Pharmacy of UFRGS first answered a questionnaire to obtain epidemiological data. Cytopathological diagnosis, Papanicolao smear, and presence of Candida spp. yeast, culture and identification and Trichomonas vaginalis, light microscopy. During the study period, 227 patients, 25.11%, aged 15 to 82 years, presented positive culture for Candida spp. Six species were identified, C. albicans (40.35%), C. glabrata (28.07%) and C. parapsilosis, 15.79%. C. krusei, C. guilliermondii and C. tropicalis totaled (15.79%). Regarding the inflammatory process, present in 49.12%, in C. krusei (75%) and C. guilliermondii (100%), and absence in C. tropicalis. The correct diagnosis and treatment of patients with vaginal candidiasis, even if not considered sexually transmitted, enables to prevent contamination through direct contact, sexual or otherwise, with other individuals and the improvement of self-esteem and quality of life. The prevalence of Candida albicans is still the main cause of vaginal candidiasis, but not neglecting the increase in the number of cases associated with non-albicans species, as well as other infectious agents such as Trichomonas vaginalis, and bacteria.

A Biochemiluminescent Sialidase Assay for Diagnosis of Bacterial Vaginosis

Bacterial vaginosis (BV) is a common condition among women of reproductive age. A sensitive, quantitative and rapid assay is needed for the diagnosis of and, particularly, therapy monitoring for BV. Bacterial sialidase appears to play an important role in bacterial biofilms on vaginal epithelium, a condition closely associated with BV. Here, we report a biochemiluminescent sialidase assay that uses a substrate derivatized with firefly luciferin. In the presence of sialidase in the reaction, the substrate is cleaved to release luciferin, which is subsequently oxidized by firefly luciferase to generate a light signal. Thus, the light signal intensity can be used to detect and measure the relative concentration of sialidase in a vaginal sample as a means of BV diagnosis. All reagents are present in a reagent bead and sample buffer, enabling essentially a one-step assay. The assay is highly sensitive and quantitative, with a sensitivity and specificity of 95.40% and 94.94%, respectively, compared to the Amsel method. Interestingly, only 27.6% of those with BV had high levels of sialidase activity with a signal to cutoff ratio of 10 or more. The assay may be used for diagnosis of BV, risk assessment of BV patients in terms of sialidase activity levels, and monitoring antibiotic therapy.

Collinsella vaginalis sp. nov. strain Marseille-P2666T, a new member of the Collinsella genus isolated from the genital tract of a patient suffering from bacterial vaginosis

A strictly anaerobic, Gram-stain-positive, non motile and non-spore-forming rod-shaped bacterium, strain Marseille-P2666T, was isolated using the culturomics approach from a vaginal sample of a French patient suffering from bacterial vaginosis. Cells were saccharolytic and were negative for catalase, oxidase, urease, nitrate reduction, indole production, hydrolysis of aesculin and gelatin. Strain Marseille-P2666T exhibited 97.04 % 16S rRNA sequence similarity to Collinsella tanakaei type strain YIT 12063T, the phylogenetically closest species with standing in nomenclature. The major fatty acids were C18:1ω9 (38 %), C16 : 0 (24 %) and C18 : 0 (19 %). The G+C content of the genome sequence of strain Marseille-P2666T is 64.6 mol%. On the basis of its phenotypic, phylogenetic and genomic features, strain Marseille-P2666T (=CSUR 2666T=DSM103342T) was classified as type strain of a novel species within the genus Collinsella for which the name Collinsella vaginalis sp. nov. is proposed.

VALIDATION OF VAGINAL SELF-SAMPLING AS AN ALTERNATIVE OPTION IN PCR BASED DETECTION OF HPV IN CERVICAL CANCER SCREENING IN BOSNIA AND HERZEGOVINA

Cervical cancer represents a serious health problem affecting women worldwide especially in developing countries due to low socioeconomic status, inadequate health-care infrastructure, weaknesses in education on this particular issue and lack of effective screening programmes. The primary aim of this study was to assess alternative screening method for the improvement of cervical cancer prevention in conditions of Bosnia and Herzegovina (B&H), which could be applicable in other developing countries as well. The study was conducted on 101 subjects who provided their self-sampled vaginal swabs and/or cervical specimens collected by their gynecologists. Universal Human Papilloma Virus (HPV) primer set optimized to detect a wide range of HPV types was used for HPV genotyping from obtained swab samples in multiplex PCR. Amplicons were analyzed in agarose gel and Agilent 2100 bioanalyzer – a platform based on microfluid technology. Inter-rater agreement kappa (MedCalc2) was used to assess concordance between results of cervical and vaginal sample analysis. Out of 39 subjects who provided their vaginal and cervical samples, results of HPV detection mismatched in 10% of the cases. Inter-rater agreement showed good strength of coincidence between the results of cervical and vaginal sample analysis (kappa=0,748, CI=95 %). We presented an alternative PCR method for the detection of HPV based on vaginal self sampling which is affordable, informative, simple and applicable with high coverage level of defined targeted population and potentially significant in the given cultural and socioeconomic context.

Complete Genome Sequence of Lactobacillus jensenii Strain SNUV360, a Probiotic for Treatment of Bacterial Vaginosis Isolated from the Vagina of a Healthy Korean Woman

ABSTRACT Lactobacillus jensenii SNUV360 is a potential probiotic strain that shows antimicrobial activity for the treatment of bacterial vaginosis. Here, we present the complete genomic sequence of L. jensenii SNUV360, isolated from a vaginal sample from a healthy Korean woman. Analysis of the sequence may provide insight into its functional activity.