School of Clinical Medicine

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The awareness and perceptions of sexually transmitted infections among students attending the University of KwaZulu-Natal.
(2021) Mthembu, Funeka Nomvula.; Abbai, Nathlee Samantha.
A high prevalence of sexually transmitted infections (STIs) have been reported among youth globally and this high prevalence calls for global efforts to improve sexual and reproductive health in this population. The prevalence of STIs in young South African women and men is 0.50% and 0.97% for Syphilis, 6.6% and 3.5% for Gonorrhoea and 14.7% and 6.0% for Chlamydia. Increased evidence on behavioural change is dependent on the comprehensive understanding and perception of one’s own risk. Updated evidence of awareness and perception of STIs in university students is needed to inform relevant sex education programmes. The purpose of this study is to assess awareness and perceptions of STIs in students enrolled at the University of KwaZulu-Natal. Methodology The study used a quantitative research approach. This study was conducted at the University of KwaZulu-Natal in Durban, South Africa. The sample consists of 142 undergraduate and postgraduate registered students between the ages of 18 and 35 years. The study used purposive sampling to obtain the sample. A self-administered survey assessing awareness and perceptions of sexual risk behaviour and STIs was administered. Data was analysed using descriptive statistics. Means and standard deviation were used for continuous variables. Analyses were stratified by gender using Chi-square tests as it was expected that there would be differences in awareness and perceptions regarding risky sexual behaviour and STIs.. Analyses were done with STATA version 15.1. Results The study found that 78% of the students were aware of STIs. There was a significant association regarding awareness of Chlamydia infections, p=0.015. Similar to the other infections, a higher proportion of males were aware of Chlamydia when compared to females (96.4% versus 82.8%, p=0.015). Similar to Chlamydia infections, there was a significant association regarding awareness of Trichomonas across the different genders (p=0.011). According to the analysis, females are exposed to awareness of STIs from a younger age when compared to their male counterparts. Most students (34.5%) had reported that they had received information on STIs from social media and from their school teachers. There was a significant difference in the responses related to same sex practices and STI risk (p=0.047). While some students had socially acceptable perceptions, there were some that were not acceptable including sexual debut (34,5%), concern about being at risk of STI (31%), condom-less sex as an STI risk (21.2%), ease of condom negotiation (41.5%), pregnancy being more risky than STIs (28.8%) and alcohol as an STI risk (28.2%). Conclusion This study had revealed the students have high awareness of STIs. Despite the high awareness, the students still have low risk perceptions especially towards condom use, alcohol consumption and age disparate relationships. These distorted attitudes will subsequently impact on the risk behaviours and further research needs to be conducted in order to fill the gap between awareness and perception. This study highlighted the clear discrepancy between the awareness of STIs and the reported perceptions of students. Future research to evaluate STI messaging and assess actual risk versus perceived risk in this population is recommended.
Diagnostic evaluation of the BD Affirm™ VPIII assay as a point-of-care test for the diagnosis of bacterial vaginosis, trichomoniasis and candidiasis in a population of pregnant women from South Africa.
(2020) Dessai, Fazana.; Sebitloane, Hannah Motshedisi.; Abbai, Nathlee Samantha.
OBJECTIVE: Untreated Sexually Transmitted Infections (STIs) and Bacterial vaginosis (BV) pose a serious health risk to mother and child. Limited data exist on the use of the BD Affirm VPIII assay as a point-of-care test. This study compared the BD Affirm VPIII assay to the BD MaxTM Vaginal assay (reference test) for the detection of BV, Trichomonas vaginalis, and Candida spp. The prevalence of single and co-infections are also reported here. METHODS: The study enrolled 273 pregnant women from King Edward VIII hospital in Durban. Socio-demographic, sexual behaviour and clinical data were collected from all consenting women. The women provided two self-collected vaginal swabs for testing. The swabs were tested using the BD Affirm VPIII assay and the BD MaxTM Vaginal assay. The prevalence of BV, trichomoniasis and candidiasis was calculated as the percentage of women who tested positive for BV, T.vaginalis and Candida infection and 95% confidence intervals (CIs) were calculated for these percentages using the formulas for calculating CIs for proportions. The number of co-infections was calculated using chi-square analysis. The diagnostic accuracy of the BD AffirmTM VPIII assay compared to the BD Max assay was assessed through the calculation of sensitivity, specificity, Negative Predictive Value (NPV) and Positive Predictive Value (PPV) and their respective 95% confidence intervals. RESULTS: In this study population, 85% of the participants were unmarried; however, 84% reported having a regular partner, and 96.3% did not use a condom regularly. The prevalence of Bacterial Vaginosis, Candidiasis and Trichomoniasis was 49.4%, 57.2% and 10.3%, respectively. A large proportion of women (78.8%) in this study did not have a discharge despite being positive for one or more pathogens. The BD AffirmTM VPIII assay showed a moderate sensitivity (79.8%) and specificity (80.3%) for diagnosing BV in all participants. The assay had an excellent specificity for Candida and T. vaginalis of 97.4% and 100.0%; respectively, however, it exhibited poor sensitivities of 52.9% and 42.4%, respectively. CONCLUSION: Our findings show a higher prevalence of Bacterial Vaginosis in antenatal attendees than previously reported, while the prevalence of Candidiasis and Trichomoniasis was in keeping with previous reports. The high number of asymptomatic infections detected is of concern and indicates the need for the re-evaluation of the syndromic management approach, especially in the antenatal population. The BD AffirmTM VPIII assay was found to be unsuitable as a screening test for vaginal infections in pregnancy. The assay performed better as a confirmatory test and may serve useful if used in conjunction with other clinical parameters such as vaginal pH.
Genetic diversity of Gardnerella vaginalis in pregnant women diagnosed with intermediate and positive bacterial vaginosis.
(2019) Nzimande, Silondiwe Philiswa.; Abbai, Nathlee Samantha.
Bacterial vaginosis (BV) is the main cause of abnormal vaginal discharge in women of reproductive age. Gardnerella vaginalis, has been detected in almost all women with BV. However, there is limited information on the genetic diversity of G. vaginalis isolated from BV intermediate and positive cases. In this study we investigated the genetic diversity of G. vaginalis strains from South African pregnant women. Vaginal swabs were characterized by the Nugent method. A total of n= 87 samples were included in the genetic analysis, (n=50 BV positive) and (n=37 BV intermediate). The presence of G. vaginalis was detected by PCR using bacterium specific 16S rRNA primers. All PCR positive amplicons were sequenced by the Sanger method and the edited sequence data was used for the phylogenetic analysis using the PHYLIP software. The sialidase A gene was amplified by PCR using specific primers and the copy numbers of sialidase A gene was quantified by droplet digital PCR. To assess the diversity of the sialidase A gene, Sanger sequencing was performed. The 16S rRNA gene from G. vaginalis was amplified in all BV positive and BV intermediate samples. All PCR amplicons were successfully sequenced and the nucleotide BLAST results revealed 100% identify to G. vaginalis. The phylogenetic analysis revealed that there is no diversity in G. vaginalis present in BV positive and intermediate cases. The phylogenetic tree of sialidase A sequences from intermediate and positive BV cases revealed two major clades which showed differences related to sialidase A copy number. Quantification of sialidase A showed that the average number of copies per cell was much higher in the BV positive group compared to the intermediate group. Some of the intermediate cases showed high copy numbers for the virulence gene and clustered with the BV positive cases. In the present study the 16S rRNA sequences of the G. vaginalis from BV intermediate and positive women showed that there is no genetic diversity in G. vaginalis detected in BV positive and intermediate samples. The phylogenetic tree of sialidase A gene sequences of intermediate and positive BV revealed two major clades which showed differences related to sialidase A copy number. This data was previously lacking in our setting, especially in a pregnant population. We further demonstrate for the first time that the genetic information present within the sialidase A gene has a direct influence on BV status.
Genotypic and phenotypic analysis of Neisseria gonorrhoeae for the identification of resistance to various antibiotics in pregnant women attending antenatal care at the King Edward VIII Hospital in Durban, KwaZulu-Natal, South Africa.
(2021) Oree, Glynis.; Abbai, Nathlee Samantha.; Ramsuran, Veron.
Introduction: Worldwide antimicrobial resistance (AMR) is making the clinical management of sexually transmitted infections (STIs) increasingly challenging with a particular emphasis on the emergence of antibiotic resistant strains of Neisseria gonorrhoeae (N. gonorrhoeae). In the current study, the detection and emerging patterns of drug resistant clinical isolates of N. gonorrhoeae to previous and current antibiotics to treat cervicitis pathogens as per syndromic management guidelines was investigated. Methodology: This cross-sectional study was conducted at King Edward VIII Hospital and included 307 antenatal attendees. Two endocervical swabs were collected from each enrolled woman. Each enrolled women also provided data on socio-demographic, behavioural and clinical factors. The first swab was placed in Amies Charcoal media immediately after collection. This swab was used to confirm the identification of N. gonorrhoeae clinical isolates using culture based assays. Culture confirmed isolates were grown in Mueller Hinton Broth and were each adjusted to have a bacterial suspension turbidity of a 0.5 McFarland Standard. These isolates were then subjected to antibiotic susceptibility testing to ceftriaxone, tetracycline, spectinomycin, azithromycin, ciprofloxacin, penicillin G and cefixime using the Etest™ method. The second swab was processed for molecular based assays. Extracted DNA from the second swab was subjected to the TaqMan quantitative Polymerase Chain Reaction (qPCR) assay, an in-house 16S ribosomal RNA (rRNA) PCR and PCR detection of the opacity (opa) gene. DNA extracted from the endocervical swabs and cultured isolates were used for the detection of specific targets (genes/plasmids/mutations) associated with resistance to penicillin, tetracycline, ciprofloxacin, spectinomycin, cefixime, azithromycin and ceftriaxone. All statistical analysis performed in this study was conducted in RS Studio. Results: The prevalence of N. gonorrhoeae was 7.8% (24/307) when detected by the TaqMan qPCR assay and 1.9% (6/307) by culture. When compared to culture, PCR for the opa gene and PCR for the 16S rRNA, the TaqMan qPCR assay was a more superior assay demonstrating a diagnostic accuracy of 94.5%. Susceptibility testing of the six isolates obtained after culturing showed resistant phenotypes for penicillin G (12 - 64 mg/L), tetracycline (1.9 - 32 mg/L) and ciprofloxacin (1.16 - 3 mg/L). Isolates displayed either dual or triple resistance to these 3 antibiotics. However, all isolates showed susceptibility to spectinomycin (>64mg/L), azithromycin (1mg/L), ceftriaxone (>0.125 mg/L) and cefixime (>0.125 mg/L). This study also detected the resistance determinants associated with penicillin, tetracycline, ciprofloxacin, spectinomycin, cefixime, ceftriaxone and azithromycin from the molecular level using the primary endocervical swab sample. Gene mutations and plasmids associated with resistance to tetracycline (tetM gene carried on a plasmid), penicillin G (penicillinase producing plasmid) and ciprofloxacin (Ser-91 mutation) were detected confirming the results obtained with the susceptibility assays. Resistance mutations associated with the remaining antibiotics were not detected. There was a 100% correlation of cultured isolates and endocervical swabs for detecting the specific AMR determinants conferring resistance to tetracycline, penicillin G, and ciprofloxacin. Conclusion: The TaqMan qPCR assay has the ability to serve as a future diagnostic assay for the detection of N. gonorrhoeae. Despite the lack of resistance to spectinomycin, cephalosporins and azithromycin in our study population, continuous surveillance for emerging patterns of resistance to these antibiotics is still required since they form part of the current South African treatment guidelines. The detection of resistance determinants from the molecular level without the need for culture may prove to be more feasible for future epidemiological investigations focused on tracking antimicrobial susceptibility or resistance patterns in N. gonorrhoeae.
Genotyping of gardnerella vaginalis from pregnant women in Durban by amplified ribosomal DNA restriction analysis.
(2020) Pillay, Kayla.; Abbai, Nathlee Samantha.; Naicker, Meleshni.
Gardnerella vaginalis is one of the most frequently isolated microorganisms associated with bacterial vaginosis (BV). However, limited information concerning the genetic diversity of G. vaginalis isolated from BV positive and intermediate cases, has been documented. This study investigated the diversity of G. vaginalis in pregnant women, a currently under-researched area in South Africa. The study population included pregnant women recruited from a public hospital in Durban, South Africa. The women provided 2 self-collected vaginal swabs for microscopy and the genotyping assays. The BV status of the women was determined using Nugent scoring. A total sample of n=137 specimens was selected for analysis. The 16S ribosomal ribonucleic acid (rRNA) gene of G. vaginalis was used for the genotyping assays. The 16S rRNA gene polymerase chain reaction products were digested with TaqI to generate genotyping profiles and genotypic subtypes were determined by correlating BamHI and HindIII digestion profiles. Phylogenetic analysis was performed on the 16S rRNA gene sequences. The data analysis was performed in R Statistical Computing software, version 3.6.2. Restriction digestion with TaqI revealed the presence of two different genotypes i.e. GT1 and GT2. Within both BV positive and intermediate sample groups, GT1 was the most prevalent genotype (54%). Overall, 4 subtypes (1, 2B, 2AB and C) were shown to be present in the sample population. The most prevalent subtype was 2B (15/37, 40.5%), followed by subtypes 1 (11/37, 29.7%), 2C (4/37, 10.8%) and 2AB (4/37, 10.8%). The phylogenetic analysis of the 16S rRNA genes showed the presence of 5 clusters. The tree displayed clusters which contained groups of specimens from the same BV group with different genotypes and subtypes present. There were also clusters which contained specimens from across the BV groups carrying the same genotype and subtype. Finally, the study did not find a significant association (p>0.05) between reported symptoms of discharge and genotype harboured. This study provides the first report on the diversity of G. vaginalis in South African pregnant women. Diversity assessments of G. vaginalis with respect to genotypes and subtypes may aid in a greater understanding on the pathogenesis of this microorganism.
A novel study to detect and quantify microorganisms associated with bacterial vaginosis from urine samples.
(2019) Naicker, Deshanta.; Abbai, Nathlee Samantha.; Naicker, Meleshni.
Bacterial vaginosis (BV) is the most common vaginal condition found in women of reproductive age. The lack of published data on the detection of BV-associated pathogens from urine, a non-invasive sample, lends novelty to the present study. This study aimed to detect and quantify Gardnerella vaginalis, Prevotella bivia, Atopobium vaginae and Lactobacilllus crispatus from urine, as an alternative non-invasive method to vaginal swabs from pregnant women using droplet digital PCR (ddPCR). A total of n=100 DNA samples (50 paired urine and swabs) were tested. The samples were stratified as BV negative and positive using the BD MAX Vaginal panel assay (Becton Dickinson). Total DNA was extracted from urine (10 ml) and swabs (1 ml) using the PureLink Microbiome Kit (ThermoFisher Scientific). The concentration of extracted DNA for urine and swab samples was determined using the Nanodrop Spectrophotometer (ThermoScientific). Droplet digital PCR was used to determine the absolute quantification of the pathogens using commercially available primer and probe sets. G. vaginalis was observed as the most abundant microorganism, followed by A. vaginae and P. bivia in the BV positive samples. When comparing abundance of microorganisms across urine and swab, it was shown that there was no significant difference across both sample types in the BV negative group. A significant difference in the BV positive group (p=0.004) was only observed for A. vaginae. Good correlation between the urine and swab was observed for G. vaginalis (R=0.63, p<0.0001), L. crispatus (R=0.71, p<0.0001) and P. bivia (R=0.50, p<0.0001). However, a weak correlation across both sample types was observed for A. vaginae (R= 0.21, p=0.001). We observed that urine has the potential to serve as an alternative sample collection method to detect BV-associated bacteria. In addition, the data generated in this study provides a basis for the development of ddPCR as a diagnostic tool for BV.

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