bluuebunny/crossref_metadata_2025_split huggingface dataset:
P4-S1.08 Performance comparison ofNeisseria gonorrhoeae(NG) sequence typing methods NG-MAST and NG-MLVA on a defined panel of isolates
Authors: Heymans R , Golparian D , Bruisten S , Unemo M 
Published: July 2011

BackgroundThe increase inN gonorrhoeae(NG) prevalence since the mid-1990s, increasing antibiotic resistance, and the absence of an effective vaccine, demand more efficient national and international surveillance of the NG epidemiology. Accordingly, objective, robust and discriminatory genotyping is a crucial tool. The performance characteristics of the internationally frequently used NG multiantigen sequence typing (NG-MAST) and the recently developed multi-locus variable-number tandem repeat analysis (NG-MLVA), were compared.MethodsFrom 2002 to 2003, NG isolates (n=330) were obtained from multiple anatomical locations from the same patient (n=118) and from self reported sexual contacts (54 couples). These isolates, were genotyped using NG-MAST, by sequencing of more polymorphic segments of porB and tbpB, and by NG-MLVA, which examines polymorphisms in five different tandem repeat loci.ResultsFrom the 118 patients with multiple isolates per visit, 97 (82.2%) had identical NG-MAST STs. Of these 97, 68 patients (57.6%) had isolates with identical MLVA types, 22 patients (18.6%) had single-locus variants (SLVs), six patients (5.1%) had double-locus variants (DLVs), and one patient (0.8%) had a multilocus variant. Of the isolates of 13 patients (11.0%) with non-identical NG-MAST STs, nine patients had identical MLVA types, three patients had SLVs, and one patient had a DLV. For the isolates of the remaining eight patients (6.8%), both typing methods identified divergent genotypes. So for 76 patients (64%) fully concordant clustering was obtained. Considering a SLV as an identical MLVA type the concordance between the two techniques increased to 83%. From 44 of the 54 couples (81.5%), the isolates of the patient and the contact had identical NG-MAST STs, while there were 34 MLVA couples (63.0%) with identical MLVA types, seven couples (13.0%) with SLVs, and three couples (5.6%) with DLVs. The isolates of 6 couples (11.1%) had identical MLVA types with different NG-MAST STs. In the isolates of four couples (7.4%) both typing methods identified divergent genotypes. So for 45 couples (83%) the two typing techniques were concordant.ConclusionsFor efficient international surveillance of gonorrhoea transmission, objective, highly discriminatory, but stable genotyping is essential. Both NG-MAST and NG-MLVA displayed a high resolution and are sequence-based portable genotyping methods. They can either be used singly or in combination, depending on epidemiological questions.
P4-S3.01 Macrolide-resistance testing and molecular subtyping of Treponema pallidum strains from South Africa
Authors: Muller E , Paz-Bailey G , Lewis D 
Published: July 2011

ObjectivesTo determine whether the main 23S rRNA mutation (A2058G) that confers macrolide resistance inTreponema pallidumis present among DNA obtained from syphilitic ulcers in South Africa and to determine the strain subtype distribution using molecular methods.MethodsDNA was re-extracted from sixty ulcer swabs, previously shown to containT pallidumDNA by in-house real-time multiplex PCR assay, using a MagNA Pure Compact DNA extraction system (Roche, Germany). The genital ulcer specimens were collected in South Africa between 2005 and 2009 during either national microbiological surveillance activities or as part of large episodic acyclovir treatment trial (HSV4294). The re-extracted DNA was screened for the A2058G point mutation in the peptidyltransferase region of the 23S rRNA subunit using a rapid PCR-based restriction digest assay. Subtyping ofT pallidum, using a previously-published technique, was based on the molecular characterisation of two variable treponemal repeat genes, arp and tpr.ResultsMacrolide resistance profiles and the subtype distribution for all 60 DNA samples were obtained. The mean age of all participants was 27.4 years (range 18–43; SD 5.24) of which 55 (92%) were male and 5 (8%) were female. None of the samples analysed contained the 23S rRNA gene point mutation that confers macrolide resistance. A total of eight arp repeat sizes, 8 RFLP patterns and a combined total of 17 subtypes were identified in this study population. The most common subtypes were 14d (43%), followed by 17d (13%), 14b (7%), 22b (5%) and 23b (5%).ConclusionsThis is the first South African study to examine both macrolide-resistance profiles and the subtype distribution of syphilis strains using molecular techniques. Macrolide-resistantT pallidumstrains appear to be uncommon in South Africa compared to more developed countries. Our results indicated subtype 14d to be the predominant circulatingT pallidumstrain in South Africa and there seem to be a high degree of genetic heterogeneity within this population.
P4-S3.05 Clinical titres and stability ofTrichomonas vaginalisRNA in urine specimens
Authors: Catania M , Bennett C , Pae A , Weinbaum B , Getman D 
Published: July 2011

BackgroundThis study estimated the range of concentrations ofTrichomonas vaginalis(TV) present in naturally infected female urine specimens and evaluated the ability of the APTIMATrichomonas vaginalis(ATV, Gen-Probe Incorporated) Assay to detect a clinically-relevant amount of TV cells spiked into TV-negative male and female urine samples and stored at various temperatures.MethodsFemale urine samples collected as part of a prospective, multicenter US clinical trial were tested with the ATV Assay, a nucleic acid amplification test for the diagnosis of TV infection in asymptomatic and symptomatic women. To determine TV cell titres, serial dilutions of TV-positive urine samples were tested and results were compared to results from serial dilutions of a laboratory culture of TV with a known cell titre. To assess the stability of TV cells in urine samples, 10 male and 10 female urine samples from non-infected volunteer donors were spiked with a cultured strain of TV, stored at 4°C, 20°C and 30°C, and tested daily for up to 14 days with the ATV assay.ResultsOf 39 randomly selected TV-positive female urine samples, the median titre in unprocessed samples was 311 cells/ml (mean=2040 cells/ml; SD=4765 cells/ml), with a range of 2–28 430 cells/ml. Of these 39, 87.2% (34/39) had a TV cell titre of ≥20 cells/ml in neat urine see Abstract P4-S3.05 figure 1. To assess stability of TV cells in urine, freshly collected male and female urine samples from volunteer donors were spiked with TV to 20 cells/ml (4 cells/reaction in the ATV assay), stored at various temperatures, and then tested with the ATV assay. For samples stored at 4°C, the ATV assay was 100% reactive for both male and female samples after 14 days of storage. For samples stored at 20°C, male samples were 100% reactive after 7 days storage, while female samples were 100% reactive after 6 days of storage. For samples stored at 30°C, 100% reactivity was obtained at 2 days storage for male samples and 4 days storage for female samples.Abstract P4-S3.05 Figure 1Two-phylotype population structure of globalTrichomonas vaginalisisolates.ConclusionsThis study shows urine samples from women infected with TV have a wide range of cell titres, with an average of ∼2000 TV cells/ml. TV can be detected for 14 days when stored refrigerated or for about 1 week at 20°C. The use of sensitive, automated molecular tests such as the ATV assay for testing urine samples should facilitate screening for TV.
P4-S3.06 Study on early infection of the low genital tract of female mice by different serotypesUreaplasma urealyticum
Authors: Yang B 
Published: July 2011

ObjectiveTo observe the early inflammatory response by animal model with low genital tract Uu infection for investigating the relationship between the serovars and their pathogenicity.MethodsFifty 8–10-week-old, female BALB/c mice were randomly divided into five groups, namely, control group, serovar 1 group, serovar 3 group, serovar 4 group, and serovar 8 group, and then treated respectively with vaginal inoculation of liquid medium and Uu inoculae of serovars 1, 3, 4 and 8 after subcutaneous injection of oestradiol benzoate. The vulval symptoms and vaginal and cervical swabs were observed weekly after Uu inoculation. Four weeks later, all mice were sacrificed and their cervixes and vaginae were removed for histopathology.ResultsThere was a significant difference in the inflammation severity of cervical mucosa (p<0.001–0.05) except vaginal mucosa among all groups. The inflammation severity of cervical mucosa in serovar 4 and 8 infection was superior to that in serovar 1 infection (p<0.01–0.05), but there was no statistical difference among other groups (p>0.05).ConclusionsThe results suggest that Uu could colonise and infect the cervical columnar epithelium other than vaginal squamous epithelium. Uu-induced inflammation response is different among its serovars. Serovars 4 and 8 are more virulent compared with biovar 1 such as serovar 1.
P4-S3.07 Population genomics ofTrichomonas vaginalisreveals a globally distributed two-phylotype population structure
Authors: Conrad M , Gorman A , Schilinger J , Sullivan S , Upcroft J , Gonzalez J , Fiori P L , Arroyo R , Ravel J , Secor W E , Carlton J 
Published: July 2011

ObjectiveTrichomonas vaginalis, the causative agent of human trichomoniasis, is the most prevalent non-viral sexually transmitted infection and has been associated with increased risk of HIV transmission, making detection and treatment a global health priority. In this study, we evaluate the population genomics of globally distributed clinical isolates to characterise genetic diversity and identify population structure.MethodsWe use a panel of 21 microsatellite and three single copy gene markers to evaluate the population genomics of 18 clinical isolates collected from female patients attending New York City STD clinics in 2008, as well as 177 extant isolates collected from the USA, Mexico, Chile, Italy, Southern Africa, Papua New Guinea, and Australia. We use a panel of population genetic tools including Arlequin 3.11 and FSTAT to calculate expected heterozygosity (HE) and population differentiation (FST) statistics. To infer population structure, we use STRUCTURE 2.2, Network 4.516, and SeaView 4.2.4. We test for significance in clinical and demographic variables (χ2and t-tests) using JMP Genomics 5.0.ResultsWe detect significant genetic diversity within the species (HE=0.66) that is observed across global regions (range 0.52–0.67), and find that a two-phylotype population structure is maintained globally with few instances in population differentiation defined by geographical origin. This two-phylotype structure is further supported by minimum spanning networks, hierarchical clustering and phylogenies inferred from three single copy genes. These two phylotypes appear at nearly equal frequencies globally. Defining characteristics of the phylotypes include significantly different prevalence ofT vaginalisvirus infection (3% in phylotype 1 and 73% of phylotype 2 (p<0.0001, χ2)) see Abstract P4-S3.07 figure 1 and a significantly higher minimum inhibitory concentration of metronidazole in phylotype 1 (p=0.0024, χ2Wilcoxon Rank Sums).Abstract P4-S3.07 Figure 1TVV infection status by phylotype.ConclusionsWe detect clear population structure and high levels of genetic diversity from globalT vaginalisisolates. This preliminary data suggests that phylotypes differ phenotypically. An accurate survey of genetic diversity and population structure will facilitate responsible selection of drug and/or vaccine targets for future treatments, and will enable better understanding of virulence factors contributing to the wide range of severity of symptoms associated with trichomoniasis.
P4-S3.10 ATrichomonas vaginalishuman vaccine candidate: Freund's adjuvant vs aluminium hydroxide adjuvant in a BALB/c mouse vaginal infection model
Authors: Smith J , Garber G 
Published: July 2011

BackgroundTrichomonas vaginalisis an underreported STD known to increase HIV transmission. Development of a vaccine againstT vaginaliscould greatly affect transmission ofT vaginalisand HIV as well as reducing their global prevalence. Herein, we aim to investigate the feasibility of developing aT vaginalisvaccine using the human-safe adjuvant Alhydrogel compared against an already established Freund's adjuvanted vaccine in a BALB/c vaginal infection mouse model.MethodsA prime-boost vaccination schedule was employed with live, whole cellT vaginalis(1×107 Tv/ml) is injected in 2–200 ul aliquots 4 weeks apart and prior to vaginal infection challenge. Either Freund's, 0.5 mg Al/ml Alyhydrogel, or 0.75 mg Al/ml Alhydrogel were applied as adjuvants. Additionally, 0.75 mg Al/ml Alhydrogel sham and non-vaccinated groups serve as controls. The BALB/c vaginal environment was modified to mimic humans by adjustment of vaginal flora and estrogenisation prior to vaginal infection. The total IgG, IgG1 and IgG2a levels in serum were tested 3 weeks post each vaccine injection, then 2 and 4 weeks post-vaginal infection by ELISA. Groups were compared using Tukey's MCT (Post-Hoc one-way ANOVA; α<0.05).ResultsAlhydrogel adjuvanted groups showed comparable total IgG and IgG1 levels to Freund's adjuvanted group. These levels for all adjuvanted groups were significantly different from control groups at all time points following boost-vaccination. IgG2a levels were not as consistent with large SD seen within Freund's adjuvanted mice. Alhydrogel adjuvanted groups were not significantly different from control groups at any time point for IgG2a levels suggesting no induction of IgG2. Controls and sham vaccines showed no Ig response.ConclusionsOur data suggest that in line with expected Th2 skewed response from Alhydrogel adjuvant and Th1 skewed response from Freund's adjuvant there was a difference in IgG2a antibody production. Additionally, the Alhydrogel adjuvanted vaccines are otherwise similar to Freund's adjuvant in terms of total IgG and IgG1 levels suggesting the feasibility to pursue Alhydrogel as a vaccine candidate. Alhydrogel induces a significantly elevated immune response compared to natural infection antibody production (non-vaccinated control) and does not induce a nonspecific immune response.
P4-S4.02 A 22-Organism microarray approach for detecting microbiological associations with symptomatic urethritis in males
Authors: Patel S , Pond M 
Published: July 2011

BackgroundIn addition to its known microbiological aetiology, urethritis in men may be linked with other genital tract organisms, as yet unidentified in its pathogenesis. We used a microarray, with capacity to detect 22 genital tract organisms, in order to determine the association of symptomatic urethritis with infection or carriage of these organisms.Methods129 patients were asked to provide an extra first void urine specimen or give permission for their residual urine specimen submitted forChlamydia trachomatisNAAT testing to be utilised. Patients were categorised into three self-reported symptom groups: definite symptoms of urethritis (discharge and/or dysuria), category 1(C1) n=80; non-specific symptoms of urethritis (eg, minimal urethral discomfort), category 2 (C2) n=26; and asymptomatic category 3 (C3) n=23. Total urine nucleic acid was extracted and subsequently used for PCR coupled microarray analysis. Organisms were defined as present or absent using an online data analysis method. In a pre-planned analysis, the following categories were compared for prevalence of organisms: C1 vs C2 and C3 combined; C1 and C2 combined vs C3 using Fisher's exact test.ResultsOne or more organisms were detected in 74% (n=95) of patients and two or more organisms in 33% (n=42). The prevalence of organisms known to cause urethritis in this largely symptomatic cohort was: 16% (n=21), 9% (n=12) and 5% (n=6) forC trachomatis,Mycoplasma genitaliumand forNeisseria gonorrhoeaerespectively.Escherichia coliwas the most prevalent organism detected with a prevalence of 18% (n=23). The presence ofM genitaliumwas statistically associated with C1 and C1 and C2 combined (p=0.03 and 0.01 respectively). In symptomatic patients,C trachomatis,Ureaplasma urealyticum, andGardnerella vaginalisappeared to be more prevalent than in asymptomatics although not statistically significantly. Lactobacilli where detected in 1.3% and 4% of patients with C1 and C2 symptoms respectively, compared with 17% of asymptomatic patients. The absence of lactobacilli was associated with urethritis symptoms, either C1 alone or C1 and C2 combined (p=0.03 and p=0.01) respectively.ConclusionsUsing a polymicrobial microarray approach we have demonstrated that symptomatic urethritis is associated with depletion of lactobacilli. This confirms early work using urethral swabs. The temporal nature of Lactobacilli depletion in relation to the onset of symptomatic urethritis needs to be investigated further.
P4-S4.03 A low-cost microfluidics-based diagnostic test for STDs
Authors: Sia S 
Published: July 2011

BackgroundUndiagnosed and untreated STIs cause large morbidity and mortality, including birth defects and stillborn babies. Since most STI's have known treatments, the largest barriers for treating patients include high cost of transporting specimens to central labs and lack of access to diagnostic testing. We present data on a portable and low-cost microfluidics device for point-of-care diagnosis of multiple STDs (such as HIV and syphilis) in combination. In order to reduce the cost and size of the assay while maintaining high performance, we incorporated microfluidic designs such as single-use plastic microfluidic cassettes, a passive method for delivering reagents, and an amplification chemistry using gold nanoparticles.MethodsOur overall device is named mChip (mobile microfluidic chip for immunoassay on protein markers). We demonstrated an ability of mChip to simultaneously detect antibodies against HIV and syphilis in needle-pricked sample volumes. Both commercial specimens and archived specimens from Sub-Saharan Africa were used.ResultsThe test sensitivities and specificities for detection of HIV-specific antibodies and treponemal-specific antibodies matched the performances from lab-based ELISA. Compared to ELISA, our test can be performed anywhere, uses a very small volume of blood, and is about 10 times faster. Some of these results are in press inNature Medicine, 2010.ConclusionThe mChip provided excellent performance in the diagnosis of HIV using only 1 μl of unprocessed whole-blood and <15 min assay-time, and an ability to simultaneously diagnose HIV and syphilis with sensitivities and specificities equal to lab-based assays. Overall, we demonstrate an integrated strategy for miniaturising complex laboratory assays using microfluidics and nanoparticles to enable POC diagnostics and early detection of infectious diseases in remote settings.
P1-S1.53 Assessing HPV genotype prevalence in Australian women by Indigenous ethnicity pre-vaccination
Authors: Garland S M , Brotherton J M L , Stevens M I , Tabrizi S N I , Condon J , McIntyre P , Smith D 
Published: July 2011

BackgroundA government funded HPV vaccination program was implemented across Australia from April 2007. The aim of this study was to ascertain whether HPV genotype prevalence, prior to vaccination, differed significantly by Indigenous status, age group, Pap cytology status or area of residence.MethodsWe used women attending for routine Pap smear, (April 2005–February 2008), as the sampling frame with 34 sites across Australia selected to include adequate numbers of Indigenous and remote-dwelling women. The final recruitment of 2620 (mean 33, range 15–66 years) included 26% (684) indigenous. DNA extracts from Thin-Prep specimens were screened by HPV AMPLICOR (Roche) and in-house HPV PCR/ELISA, with positives genotyped by LINEAR ARRAY HPV genotyping test (Roche).ResultsThe overall prevalence of HPV infection was 39% (95% CI—36.8 to 40.6), with high risk (HR) HPV detected in 26% (95% CI—24.8 to 28.2). Single infections were detected in 17% (95% CI—15.8 to 18.7). While multiple infections were common overall at 22%, there was no difference in proportion of multiple HPV carriage between indigenous and non-indigenous (58% of HPV positive non-Indigenous women and 56% of HPV positive Indigenous women had multiple types detected). As with single infections, multiple type infections were less prevalent with increasing age. The six most common genotypes were—HPV 16 (8.3%), 51 (5.1%), 53 (4.7%), 62 (4.3%), 89 (3.9%) and 52 (3.8%). Age-specific HPV prevalence rates were similar for Indigenous and non-Indigenous women aged ≤30, but higher for Indigenous women aged 31–40, particularly for non-vaccine targeted HR-HPV genotypes. By HPV clades for this age group, indigenous women were significantly more likely to have α 7 HPV ((45, 39, 59, 68 or 70 without 18) OR 1.9 (1.1 to 3.3) p=0.03) or α 5 group (HPV51, 26, 69, 82) with OR 2.1 (1.1 to 4.3) p=0.02). There was no significant association between Indigenous status and detection of HPV from the α 9 clade (31, 33, 35, 52, 58, 67), with or without HPV 16. Overall, HR-HPV prevalence increased from 21% in women with normal cytology, to 81% in those with high-grade lesions/cancer.ConclusionsCross-sectional prevalence of HR-HPV was high in Australian women, with vaccine preventable genotypes observed in 13% of all women (25% in <25 year olds). Vaccination should significantly reduce vaccine related HPV infection and disease in Australian women, irrespective of indigenous status or area of residence.
P5-S1.04 The impact of pelvic inflammatory disease on sexual, reproductive and psychological health
Authors: Hocking J , Newton D , Bayly C , Fairley C , Chen M , Williams H , Keogh L , Temple-Smith M , McNamee K , Fisher J , Hsueh A , Hocking J 
Published: July 2011

Background and AimPelvic inflammatory disease (PID) is a condition commonly attributable to chlamydia infection. This qualitative study sought to explore the sexual, reproductive and psychological impact of this condition.MethodsIn depth, semi-structured interviews were conducted with 23 women diagnosed with PID. Both symptomatic and asymptomatic women were recruited from primary and tertiary healthcare services. Interviews were conducted with women from 2 to 12 months post-diagnosis in order to explore short and longer term psychological responses and experiences. A brief, self-report questionnaire containing demographic items was also completed by all women. Interview analysis was conducted using an inductive, thematic approach.ResultsNearly all women experienced some form of distress when they received their diagnosis, and the emotional impact of their diagnosis was generally prolonged. Women typically experienced emotions such as shock, sadness or anger. At the time of diagnosis, women frequently had little or no knowledge of PID and continued to experience confusion about their condition post-diagnosis. Some women reported that PID had created conflict in their intimate relationships or had impacted on the level of intimacy they shared with their partner. Almost all women reported that their sexual behaviour had changed dramatically post- diagnosis. The possibility of being infertile stood out for women as their greatest health concern and nearly all women reported changes to their health behaviours since their diagnosis.ConclusionThe findings of this study indicate that a diagnosis of PID can have significant psychosocial implications for the diagnosed individual. Recommendations for healthcare professionals are proposed.