bluuebunny/crossref_metadata_2025 huggingface dataset:
P3-S6.02 Is fourfold drop of the nontreponemal antibody titrestitres at three or 6 months after early syphilis treatment an effectiveness “criterion?”
Authors: Zhou P , Gu X , Lu H , Guan Z , Qian Y
Published: July 2011

Background

At present the criteria for treatment effective in early syphilis is the disappearance of clinical symptoms and signs and four-fold decrease of nontreponemal antibody titres within 3 to 6 months after therapy. However, how to evaluate if syphilis is cured after treatment remains difficult and controversial.

Methods

Secondary syphilis patients whose rapid plasma reagin (RPR) titres did not turn negative at least 24 months after treatment were enrolled in this study and their CSF were evaluated. The criteria for enrolment were: (1) RPR titres declined fourfold within 3 months after therapy for secondary syphilis; (2) patients denied high risk sexual behaviour following syphilis treatment; (3) RPR titre did not turn negative at least 24 months after treatment; and (4) HIV negative. The criteria for neurosyphilis were: (1) CSF leukocyte count was elevated, and/or (2) proteins were abnormal, and (3) a reactive VDRL-CSF test in the absence of substantial contamination of CSF with blood, and (4) a reactive TPPA-CSF test, and (5) a negative HIV test, and (6) with or without neurological manifestations, and (7) excluding other possible CNS infections.

Results

There were 14 male and three female patients who met the criteria for neurosyphilis. The CSF leukocyte count was elevated in 10 patients among which nine also had CSF proteins elevated. The other three patients had CSF proteins elevated only. CSF-VDRL and CSF-TPPA were reactive in all 17 patients. There were four cases presenting notable neurological and psychiatric manifestations, and other 13 had no signs and symptoms of CNS when they entered the study. The clinical symptoms and signs disappeared or improved in four patients, and CSF-WBC in those nine patients turned to normal after treatment. CSF-protein declined accordingly but did not turn to normal in four cases see Abstract P3-S6.02 table 1.

Abstract P3-S6.02 Table 1

RPR and CSF before anti-neurosyphilis treatment

No.Initial RPRRPR (3 months after treatment*)RPR (24 months after treatment*)CSF/WB (0–8)×106/lCSF/protein (150–450) mg/lCSF/VDRL11:641:81:168.1↑860↑1:421:161:41:434411:131:321:81:81.1890↑1:241:641:61:32300↑1860↑1:851:321:41:83.81050↑1:461:1281:321:648.2↑650↑1:1671:641:161:169.8↑460↑1:481:2561:321:325.8580↑1:891:641:81:826↑460↑1:4101:1281:321:328.8↑2801:8111:161:41:89↑540↑1:8121:1281:161:1617↑570↑1:4131:2561:641:6438↑590↑1:16141:1281:321:82.24301:2151:2561:321:1618.8↑800↑1:4161:641:161:83.34301:1171:161:41:432801:1*

Anti-secondary syphilis treatment.

Conclusions

A four-fold decrease in serological titres and resolution of lesions of early syphilis may not predict success. The occurrence of failure after standard therapies suggests that the current criteria for “treatment effective” are questionable. There is a need of continuing to evaluate early syphilis patients who meet the criteria for “treatment effective” and whose nontreponemal antibody titres fail to turn negative afterwards.
P3-S6.04 Use of a point of care test device to detect syphilis in a STD clinic
Authors: Barnes M , Ducan D , Agreda P , Hsieh Y H , Holden J , Rompalo A , Gaydos C
Published: July 2011

Background

Diagnosis of syphilis is problematic and an accurate rapid point of care (POC) test could be useful in busy STD clinics. There are no FDA Cleared POC tests for Syphilis serology in the USA.

Objective

To determine the performance of a new, rapid point of care (POC) test, Syph-Check, which is not yet FDA cleared, for the serological diagnosis of Treponema pallidum in female and male STD patients.

Methods

Men and women >18 yr visiting the Baltimore City Health Department STD clinic were consented to enrol in a trial to determine the accuracy of a new, innovative POC test for syphilis (Veda, manufactured in France) that used a cassette format to test syphilis serology. The Syph-Check One-Step Syphilis test is a point of care, rapid immunoassay screening test for qualitative detection of IgG and IgM antibodies toTreponema pallidumin finger stick blood, plasma, and serum. This product can be used as an initial screening test or as a confirmatory diagnostic test, but is not FDA cleared for use in screening blood or plasma donors. The assay was performed in the STD clinic, required only 20 min to perform, and required no instrumentation. RPR and TPPA tests were performed to determine the sensitivity and specificity of the Syph-Check POC test.

Results

194 men and 205 women were enrolled. Of the 399 samples tested, 33 were positive and 366 were negative by the Syph-Check. There were 14 positives and 385 negatives by RPR confirmatory testing. Overall sensitivity compared to RPR testing was 85.7% (95% CI 60.3% to 97.5%) and specificity was 94.5% (95% CI 91.9% to 96.5%). There were 32 positives and 367 negatives by TPPA confirmatory testing. Overall sensitivity compared to TPPA was 43.8% (95% CI 27.5% to 61.1%) and specificity was 94.8% (95% CI 92.2% to 96.8%).

Conclusions

The Syph-Check POC test was moderately accurate compared to the RPR test, but not as sensitive compared to TPPA. A more accurate POC test for syphilis could be useful for clinicians to test clinic patients and provide immediate screening results for syphilis to patients during an office or clinic visit.
P3-S6.05 Comparing the analytical sensitivities of six treponemal tests
Authors: Jost H , Kikkert S , Fakile Y , Castro A , Ye T , Zaidi A , Ballard R , Cox D
Published: July 2011

Background

Traditional syphilis testing consists of screening with a non-treponemal test (RPR) and confirmation with a treponemal test (TP-PA, FTA-ABS, EIA, CIA, etc). Recently, that testing algorithm has been reversed due to efforts to reduce labour costs and the availability of automatable tests (EIA, CIA). Large numbers of discordant test results (treponemal +, non-treponemal−) can be obtained using the reverse algorithm and can be due to (1) treated cases of syphilis, (2) a false-positive treponemal test, or (3) a case of early primary syphilis that has yet to seroconvert. Those sera need to be confirmed with a second treponemal test to eliminate false positive specimens. The dilemma arises which treponemal test is best suited for confirmation, and secondly what are the relative analytical sensitivities of available treponemal tests used for both screening and confirmation.

Methods

Two hundred randomised TP-PA positive, cleanascite treated samples (GADPH) were serially diluted with normal human sera to determine the analytical endpoint and sensitivity of six commonly used treponemal tests (TP-PA, FTA-ABS, TrepSure, TrepChek, TrepID, and LIAISON). All dilutions were treated as neat sera in each test, and the tests were performed according to the manufacturer's instructions.

Results

The FTA-ABS was used as the reference since it has been considered the Gold Standard" among treponemal tests. The analytical sensitivity of the six tests fell into three statistically different groups (from lowest to highest): (1) the FTA-ABS, the TP-PA, and the TrepChek, (2) the LIAISON and the TrepID, and (3) the TrepSure. In addition, there were 24 sera that were found non-reactive by FTA-ABS and nine found non-reactive by TrepChek that were reactive by at least two other treponemal tests.

Conclusions

These results highlight significant differences in the analytical sensitivity of various treponemal tests and could explain some discordant results between treponemal tests used to confirm screening EIAs and CIAs. First, the FTA-ABS should no longer be considered the “Gold Standard”. Second, low titre sera could appear non-reactive if relatively insensitive tests are used for the either screening or confirmatory tests. Third, the question arises, “Should initial screening tests be confirmed by treponemal tests with lower analytical sensitivity?” Future patient studies could verify these results, so that a more accurate standard protocol for syphilis testing can be established.
P3-S6.08 Detection of the 23S rRNA point mutations (A2058G and A2059G) associated with azithromycin resistance in treponema pallidum using a TaqMan-based real-time Triplex-PCR assay
Authors: Chen C Y , Chi K H , Pillay A , Nachamkin E , Ballard R
Published: July 2011

Background

To develop a TaqMan-based real-time allelic discrimination assay for the simultaneous detection of two point mutations (A2058G and A2059G) in the 23S rRNA gene ofT pallidumthat have been associated with azithromycin treatment failures.

Methods

Initially, two TaqMan-based real-time duplex PCR assays were used to detect the A2058G and A2059G point mutations within the 23S rRNA gene ofT pallidum. Genotyping results from these assays were then compared to a previously described real-time PCR assay using fluorescence resonance energy transfer (FRET) probes and melting curve analysis that is specific for the detection of the A2058G point mutation. Subsequently, a real-time triplex PCR was developed to distinguish the A2058G from the A2059G point mutation in a single assay and the results were confirmed by pyrosequencing.

Results

Sixteen of 67 (23.9%)T pallidum-positive specimens collected from patients with genital ulcer disease in the US were found to have the A2059G point mutation. These strains were previously characterised as having azithromycin-susceptible genotypes (no point mutations in the 23S rRNA gene). The A2059G mutation was confirmed by a real-time duplex PCR assay containing the TaqMan probe specific for the mutation and by pyrosequencing. None of theT pallidumstrains examined to date had both point mutations. The real-time triplex PCR assay was able to differentiate azithromycin-susceptible genotypes from resistant genotypes containing either the A2058G or A2059G point mutation in a single assay. The limit of detection of the assay was 1–10 copies using 23S rRNA gene fragments that were cloned into a plasmid.

Conclusions

The previously reported real-time PCR detection platforms were designed to detect only the A2058G point mutation and were unable to differentiateT pallidumstrains with susceptible genotypes from resistant genotypes with the A2059G mutation. The new TaqMan-based real-time allelic discrimination assay offers an alternative to the previously described PCR/RFLP method to rapidly detect both point mutations associated with azithromycin resistance inT pallidum. The prevalence of T. pallidum strains that harbour point mutations in the 23S rRNA gene associated with resistance to azithromycin might be much higher than previously estimated.
P3-S6.10 Analysis of systemic and cutaneous immune responses helps explain the duality of immune evasion and recognition in secondary syphilis
Authors: Cruz A , Ramirez L
Published: July 2011

Background

Despite the robust adaptive immune responses associated with secondary syphilis (SS), which include the presence of high titres of anti-T pallidum(Tp) antibodies with opsonizing activity, it may take months to years for host defenses to gain control of the infection. Tp's unusual outer membrane architecture is believed to underlie this duality of immune evasion and recognition. To better understand the dynamics of immune evasion and Tp clearance in early syphilis, we studied immune responses of SS patients in peripheral blood (PB) and skin in relation to Tp burdens.

Methods

27 HIV(-) SS patients and 26 healthy controls were enrolled. Circulating monocytes, dendritic cells (DCs) and natural killer (NK) cells were characterised by flow cytometry and qRT-PCR. SS skin biopsies were analysed by immunohistochemistry (IHC) and microarray. Ex vivo opsonophagocytosis assays were performed with live Tp (Nichols) and purified human monocytes.

Results

Despite the demonstrated presence of Tp in blood, circulating monocytes exhibited only mild activation by flow cytometry without detectable cytokine production by qRT-PCR. We also observed decreased numbers of circulating IFNg-producing and cytotoxic NK cells along with an emergent CD16+/CD56- NK-cell population. In contrast, skin lesions, which contained abundant Tp by IHC and PCR, contained transcripts for a variety of pro-inflammatory cytokines (IFNg, TNFa, IL1b), monocyte chemotactic factors (CCL2, CCL5 and CXCL10) and macrophage and DC surface activation markers (CD80, CD86). Transcripts for genes associated with Fc-mediated phagocytosis (FcgRI, FcgR3), endosomal TLRs (TLR7-9), and cytotoxic T cells (CD8, granzyme, perforin) also were detected in lesional skin. IHC corroborated the presence of macrophages and CD4 and CD8 T cells, but not NK cells, in the biopsies. Patient sera promoted Tp uptake and monocyte activation, although substantial proportions of Tp were capable of evading phagocytosis.

Conclusions

Our results support a model in which the duality of immune evasion/recognition which occurs in SS reflects the relative burdens of Tp in skin and blood as well the presence of Tp populations with differential capacity for binding opsonic antibodies. Macrophage activation due to opsonophagocytosis of a subpopulation of Tp drives the recruitment to skin of immune cells that slowly clear infection. Results also suggest that cytotoxic cellular responses may contribute to elimination of Tp.
P3-S7.01 Evaluation of a simple point-of-care rapid test for detecting Trichomonas vaginalis among women in Mysore, India
Authors: Rao K , Madhivanan P , Klausner J , Trammell S , Kotian S , Ravi K , Krupp K , Srinivas V
Published: July 2011

Background

Trichomonas vaginalisis one of the easily treatable sexually transmitted infections in the world. Current methods used to diagnose trichomoniasis rely heavily on training and experience of the technician and have low performance reliability. These methods are unsuitable for settings where accessibility to a continuous electrical source may be a challenge. We examined the performance of OSOM Trichomonas rapid point-of-care test (POC) as compared to the gold standard (culture) and the routine method of diagnosis in clinical laboratories (wet-mount microscopy) in Mysore, India.

Methods

Sexually active women over age 18 and seeking care at Prerana Reproductive Health Clinic were enrolled into the study from July 2009 to August 2010. Clinician- collected vaginal swabs were evaluated for trichomonads using wet-mount microscopy, InPouchTM culture, and OSOM® Trichomonas Rapid Test in a blinded manner by different investigators.

Results

Of the 417 women enrolled, the prevalence of Trichomoniasis diagnosed by culture was 16.3% (95% CI 12.9% to 20.3%). As compared to culture, the sensitivity, specificity, positive predictive value and negative predictive value for wet mount microscopy was 82.4%, 98.9%, 93.3%, 96.6%; for OSOM rapid test it was 88.2%, 99.4%, 96.8%, 97.8% respectively.

Conclusion

OSOM Trich rapid test had very good performance with excellent sensitivity, specificity, positive predictive value, and negative predictive value of 88.2%, 99.4%, 96.8%, and 97.8%, respectively. The implementation of OSOM Trichomonas rapid test would significantly reduce the labour and material costs. Furthermore, frequent partner reinfection as a result of wrong or missed diagnosis can be reduced. It will also reduce other complications such as pelvic inflammatory disease and susceptibility to HIV.
P3-S7.02 Performance of a point-of-care diagnostic for Bacterial Vaginosis among young reproductive age women in Mysore, India
Authors: Ravi K , Kotian S , Rao K , Selezneva J , Srinivas V , Krupp K , Madhivanan P
Published: July 2011

Background

Bacterial vaginosis (BV) remains the most common cause of abnormal vaginal discharge in Indian women of reproductive age and is associated with increased susceptibility to HIV/STI and preterm delivery. Diagnosis of BV in resource-poor settings is often overlooked; there is a need for cheap, rapid, objective point-of-care diagnostic test.

Methods

Vaginal swabs were collected from women attending a women's health clinic. Women over the age of 18 with a pH of over 4.5 were invited to participate in the study. BV was diagnosed on the basis of the Nugent score, the Amsel clinical criteria, and results of OSOM BVBlue test independently by study clinician and laboratory personnel who were blinded to the results of the other tests.

Results

From August 2009 to May 2010, 313 participants were enrolled. BV prevalence was 45.1% (95% CI 41.5% to 52.8%) according to Nugent score. When compared with the Nugent score, the sensitivity, specificity, positive predictive value, negative predictive value for Amsel clinical criteria was 61.9%, 88.3%, 81.5%, 73.7%; and for BVBlue it was 38.1%, 92.7%, 82.1%, 63.9% respectively. The performance of BVBlue can be increased if it is combined with “Whiff test where the sensitivity increases to 64.4%, sensitivity 85.6%, PPV 79.3% and NPV 73.8%”.

Conclusions

These results highlight the importance of systematic evaluation of rapid test kits as a low-cost alternative to laboratory diagnosis in resource-constrained settings. The BVBlue test is a simple, rapid, and objective test for the diagnosis of BV and has the potential to facilitate prompt diagnosis and appropriate treatment of BV in the absence of microscopy.
P3-S7.03 The prevalence of Bacterial Vaginosis among young women in urban areas in Nigeria and its major risk factors
Authors: Olawuyi O
Published: July 2011

Issue

Vaginitis is polymicrobial Sexually Transmitted Diseases (STDs), associated with some STD agents, presenting signs/symptoms such as foul smelling vaginal discharge, irritation, and itching. Among the leading infectious agents causing vaginitis among young women in urban communities in Nigeria,Gardnerella vaginalis(cause of Bacterial Vaginosis) was found to top the list. Also, unprotected sex during first sexual intercourse and multiple sexual partners are found to be the major risk factors for the wide spread of Bacterial vaginosis among the study's young women.

Method

Bacteriological and differential laboratory tests (microscopic examination, normal physiological saline wet-preparation, Potassium Hydroxide (OH) solution, whiff test, the litmus paper/PH level test, and gram staining) were carried out on High vaginal swab (HVS) specimens collected aseptically within 6 months from non-pregnant, non-menstruating young women who are not on antibiotics for a week before specimens were collected, and whose consent is sought before the study. The Study population is made up of both STD center attending and non-attending young women (mean age=25.50). In addition, a standard questionnaire (r=0.77) was administered to answer the major research/study questions on the major risk factors for Bacterial vaginosis.

Result

From the 200 HVS collected, 68 % of primary isolates were isolated:G vaginalis(51.72%),C albicans(41.92%), andT vaginalis(06.36%) respectively (p<0.0001). According to the result from the study questionnaire, among 200 respondents, 37.71% had their first sexual intercourse without any protection, 39.16% were protected during their first sexual intercourse, and 23.13% did not recollect. Also, 31.65% had engaged in sexual intercourse with more than one sexual partner before with no protection in few occasions, 37.21% had had multiple sexual partners before with protection, 31.14% declined to provide answer to the multiple sexual partner questions.

Conclusions

Bacterial vaginosis was found to be the most prevalent among young women in urban cities in Nigeria, and the major risk of contracting the infectious disease is unprotected sexual intercourse with multiple partners, and during first sexual intercourse. However, there is need for effective and efficient sex education and awareness to encourage the partners to be faithful to one another and for the promotion of safe sexual intercourse by using preventive means such as condoms.
P3-S7.05 Phenotypic characterisation of clinical trichomonas vaginalis isolates among AAWSW in sexual partnerships
Authors: Muzny C , Rivers C , Mena L , Schwebke J
Published: July 2011

Background

Trichomonas vaginalis(TV) is the most common non-viral sexually transmitted infection in the world. TV prevalence in US women is consistently highest among African Americans (AA). However, little data exists on the risk of TV infection among women who have sex with women (WSW) and the mechanism of transmission is poorly understood. The goal of this study was to evaluate the concordance of TV isolates among AAWSW involved in sexual partnerships using the random amplified polymorphonuclear DNA (RAPD) technique.

Methods

AAWSW involved in sexual partnerships and participating in a cross-sectional study of STI prevalence rates at the Mississippi State Department of Health STD clinic in Jackson, MS, were selected for this study if both women in the partnership were infected with TV. All women completed a confidential survey asking detailed sexual history questions about female and male partners during the past 12 months. The RAPD technique utilising six unique PCR primers was performed on TV clinical isolates from each sexual partnership. RAPD amplicons were collated and categorised to determine genetic similarity between isolates from paired couples.

Results

TV isolates from three paired couples were included in this study. RAPD profiling demonstrated that only one out of the three pairs of TV isolates was concordant see Abstract P3-S7.05 figure 1. Within this concordant pair, both women reported engaging in oral sex and sharing wet towels during sexual activity. One woman in this pair reported recent sex with a male partner while the other woman denied history of other sexual partners during the past 3 months and had not had sex with a male partner in 5 years. Additionally, a follow-up visit of one of the members of this concordant union demonstrated a RAPD pattern discordant with previous findings indicating that the individual's initial treatment was successful and that she had acquired a new TV infection.

Abstract P3-S7.05 Figure 1

Primer Set 1.

Conclusions

Given the phenotypic similarity of banding patterns within one AAWSW sexual partnership, female to female transmission of TV may have occurred. The frequency of TV transmission between WSW is unknown at this time; however, the use of RAPD appears to be informative for differentiating isolates of TV. A prospective study examining the epidemiology and incidence of TV infection among WSW is necessary.
P3-S7.06 Vaginal discharge in women living with HIV attending an AIDS clinic in Manaus, Brazil
Authors: Miranda A , Silva L , Braga J V , Montes R , Talhari S
Published: July 2011

Background

A vaginal discharge and/or vulvar itching and irritation usually characterise Vaginitis, and a vaginal odour might be present. The three diseases most frequently associated with vaginal discharge are BV (replacement of the normal vaginal flora by an overgrowth of anaerobic microorganisms, (Gardnerella vaginalis), trichomoniasis, and candidiasis.

Objectives

To estimate the prevalence of vaginal discharge in HIV women attending the Institute of Tropical Medicine in Manaus, Amazonas, Brazil.

Methods

A cross-sectional study performed among women attending the AIDS clinic from March to December 2010. They were invited to take pat in the study and answered an interview including demographic, behavioural and clinical data. They underwent in a gynaecological examination and it was collect vaginal samples for diagnosingTrichomonas vaginalis, Gardnerella vaginalisandCandida spp.

Results

A total of 338 women were included in the study. Median age was 32 (IQR (IQR): 27; 38) years and median of schooling nine (IQR: 4; 11) years. Prevalence rate of vaginal discharge was 45.8% (95% CI 40.5% to 51.1%). Prevalence ofTrichomonas vaginaliswas 1.2% (95% CI 0.5% to 2.4%),Gardnerella vaginalis35.8% (95% CI 30.7% to 40.9%) and Candidiasis 21.3% (95% CI 16.9% to 25.7%). Median of first sexual intercourse was 16 (IQR: 14; 17) years and 53.6% were married or reported a stable partner. Risk factors reported were: injecting drug use (1.2%), no-injecting drugs (15.2%), previous STI (32.4%), commercial sex workers (16.4%), more than one partner in the last year (12.4%) and in life (94.7%). Regarding clinical symptoms, 50,9% reported chronic pelvic pain, 53.3% vaginal discharge, 47.6% vaginal itching, 22.8% dysuria and 9.5% genital bleeding. CD4 counts were more than 500 cells/mm3in 29.4% and viral load were <1.00 copies/ml in 53.8%. A total of 53.9% of women reporting vaginal discharge had a positive test for at least one disease. In the final model of logistic regression the only variable remained was having viral load <1000 copies/ml decreased the risk of vaginal discharge.

Conclusions

Regardless the low cost and large availability of GRAM stain and cytological tests in health services for women with AIDS, there are difficulties which remain to identify interventions that refer to social, cultural and environmental influences on vaginal infections in this group.
P3-S7.07 Detection ofTrichomonas vaginalisin HIV positive women in Pretoria, South Africa
Authors: Kock M , Rukasha I , Dijkmans A , Hoosen A
Published: July 2011

Background

The aim of this study was to detectTrichomonas vaginalisinfection in HIV positive women receiving anti-retroviral therapy in Pretoria, South Africa.

Methods

Self-collected vaginal swab specimens from 95 consecutive patients attending the anti-retroviral clinic (Tshwane District Hospital) were analysed.Trichomonas vaginaliswas diagnosed by wet mount microscopy, culture using InPouch and a commercial PCR assay targeting the DNA repeat units. Trichomoniasis was diagnosed if any test was positive.

Results

Five (5.3%) of the 95 specimens were positive by wet mount microscopy, 21 (22.1%) were culture positive and 28 (29.5%) were detected by PCR. All culture and wet mount positive specimens were PCR positive. The sensitivity and specificity of wet mount microscopy compared to culture were 23.8% and 98.7% respectively. PCR detected seven additional positive specimens than culture. The specificity of PCR compared to culture was 100%, with a sensitivity of 90.5%. The prevalence ofT vaginaliswas found to be 29.5% in this study.

Conclusions

Previous studies in South Africa focused on the prevalence of trichomoniasis in pregnant women and women without HIV status and from lower socio-economic groups. This is the first report in HIV positive women receiving ARV treatment. There was a high prevalence (29.5%) ofT vaginalisin this group. This is similar to that reported from Nigeria (24.4%) and Ivory Coast (27.0%), whilst the rate reported in Congolese (18.6%) HIV positive women was lower. Using microscopy alone for the diagnosis of trichomoniasis as is the current practice in most laboratories in South Africa is inadequate and leads to missed infections.
P3-S7.11 Factors associated with persistent Bacterial Vaginosis among young reproductive age women in Mysore, India
Authors: Madhivanan P , Rathod S , Krupp K , Arun A , Reingold A , Cohen C , Klausner J
Published: July 2011

Background

Bacterial vaginosis (BV) is a common infection and has been associated with adverse health outcomes including preterm birth, pelvic inflammatory disease and acquisition of HIV and other sexually transmitted diseases. There are limited data on persistent BV in Indian women. This study examined the risk factors for persistent bacterial vaginosis in young reproductive women in India.

Study Design

Between November 2005 and January 2007, a prospective cohort study was carried out to examine the relationship of BV and HSV-2 acquisition among women in Mysore. Quarterly, data were collected on sociodemographics, risk behaviour, partner characteristics, followed by a physical examination to diagnose and treat reproductive tract infections. BV was defined using Nugent scoring of gram stained vaginal smears. Persistent BV was calculated using generalised estimating equation methods. Women gave informed consent prior to enrolment in the study.

Results

Of the 420 women for which there were data available for all three visits, 114 (27%) had two or more BV episodes. Women with a history of 2 or more BV episodes were more likely to be infected withTrichomonas vaginalis[OR 72.93, 95% CI 9.69 to 548.4] and be diagnosed with HSV-2 infection [OR 2.58, 95% CI 1.44 to 4.63] compared to women with no BV history. Women with a history of BV were also more likely to report no education, tubal ligation, being a non-Muslim, and having a sex partner who had other sex partners.

Conclusions

Young reproductive age women in India have a high persistence of BV. Although the association between BV andTrichomonas vaginalisis unclear, it seems prudent to recommend that women with BV or TV be screened for both infections.
P3-S7.13 Evaluation of the Affirm VPIII microbial identifications test for the diagnosis of vaginitis and bacterial vaginosis
Authors: Maseko D V , Radebe F M , Gumede L , Sewphershad N Bhojraj , Khumalo S , Basson I , Lewis D
Published: July 2011

Background

Vaginitis and bacterial vaginosis (BV) are the most prevalent causes of vaginal symptoms among women of childbearing age. Diagnosis is based on clinical signs and symptoms, microscopy, pH and the whiff" test, and laboratory performed tests. The Affirm VPIII Microbial Identification test (Becton Dickinson) incorporates non-amplified DNA probes forCandida spp.,Trichomonas vaginalisandGardnerella vaginalisas an indicator for BV.

Objective

To compare the Affirm VPIII Microbial Identification test for the detection ofCandida spp.,Gardnerella vaginalisandTrichomonas vaginalisto graded Gram stain for Bacterial Vaginosis and yeast as well as to an in-houseTrichomonas vaginalisPCR (Polymerase Chain Reaction).

Methods

In this study, specimens from 191 patients were evaluated. Specimens were collected consecutively from patients with vaginal discharge syndrome presenting at an STI (Sexually transmitted infection) clinic in Gauteng, South Africa between May 2010 and February 2011. Study inclusion was dependent upon the request and collection of both a vaginal swab for Affirm VP III and a second swab to make a smear for Gram staining. A cervical swab specimen was also collected for PCR at the same visit. Graded Gram stain for Bacterial vaginosis and yeast as well as an in-house real-time PCR method on the Rotorgene platform was performed in the laboratory. The sensitivity and specificity of the assay was determined using the graded Gram staining as a gold standard for BV andCandida spp.. The real-time PCR was the gold standard for theTrichomonas vaginalis(TV).

Results

Analysis of the Affirm VPIII gave a sensitivity of 98% for BV correctly categorising 80of the 82 BV positive specimens and specificity of 76% correctly identifying 38 of the 50 BV- negative specimens. The Affirm VPIII for Candida yielded a sensitivity of 86% and specificity of 95%. When the Affirm was compared to the Real-time TV PCR the sensitivity was at 45% and specificity at 99%.

Conclusion

The Affirm VPIII is an objective system which detects mixed vaginal infections and can be used in any setting. The performance characteristics of the Affirm VPIII for BV and Candida were comparable to those of other published studies. However in this study the Trichomonas vaginalis PCR was used as a gold standard therefore the sensitivity of the Affirm VPII of 45% is similar to that of culture or wet mount.
P3-S7.15 Diagnostic reliability of wet prep microscopy forT Vaginalisin women visiting a high-volume STD clinic
Authors: Fitch J , Anderson T , Thrun M , Mettenbrink C
Published: July 2011

Background

For years, wet prep microscopy has been the cornerstone of testing forT vaginalisdue to the simplicity of the test and its low cost to perform. Though still recommended as a means of diagnosis for vaginal infections, its diagnostic sensitivity (typically 50%–70%) falls short of other means of testing, including culture (approximately 80% sensitivity). This study compares the performance and costs of wet prep tests to culture in the hands of experienced laboratorians in a high volume STD clinic.

Methods

The Denver Metro Health (STD) Clinic sees over 800 patients and performs over 100 wet prep tests every month. Between 1 January 2010 and 30 September 2010, a wet prep test and aT vaginalisculture were performed on every female patient receiving a comprehensive exam, regardless of symptoms, as part of resistance testing for the STD Surveillance Network (SSuN). The results of the two tests were then compared.

Results

Wet prep tests andT vaginaliscultures were performed on 555 female patients. Of those, 51 were positive by wet prep and 62 were positive by culture see Abstract P3-S7.15 table 1. There were no wet prep false positives reported. The average material cost for each in-house wet prep was $0.32 and the cost for each culture was $1.78. In comparision, materials for PCR testing forT vaginaliscost approximately $2 per test. Staff time required to perform wet prep microscopy is less than either culture or PCR.

Abstract P3-S7.15 Table 1

Diagnostic reliability of wet prep microscopy forT Vaginalisin women visiting a high-volume STD clinic

Wet prepPositiveNegativeTotalCulture Positive511162 Negative0493493 Total51504555

Conclusions

The wet prep sensitivity rate of 82.2% when compared to culture (which, until recently has been the gold standard for trich testing) suggests that wet prep microscopy may be more useful than previously recognised in venues where clinicians and laboratorians gain experience by performing a large number of tests. Additionally, the costs, both in supply and staff expense, suggest that, in tight budgetary times, wet prep may continue to be a cornerstone ofT vaginalistesting, even with the availability of molecular amplification methods. These results, however, were limited to a single clinic during a single point in time. As next steps, wet prep testing might be compared to molecular amplification tests.
P3-S7.17 Noninvasive high resolution imaging with optical coherence tomography for vaginal product safety assessment in women
Authors: Vincent K , Stanberry L , Moench T , Breitkopf C R , Loza M , Wei J , Paull J , Grady J , Motamedi M , Rosenthal S
Published: July 2011

Background

Prevention of HIV and other STDs using vaginal microbicides must be safe. Colposcopy has not been shown to predict microbicide safety; therefore a more sensitive method is needed in safety evaluations of these vaginal products. The use of optical coherence tomography (OCT) has shown increased sensitivity to tissue injury over colposcopy in animal models. We describe a randomised double blind clinical trial using OCT to detect epithelial changes and injury related to vaginal microbicide use.

Methods

30 women aged 18–45 were randomised to use hydroxyethyl cellulose (HEC) placebo or nonoxynol-9 (N-9) vaginal gel twice daily for 5.5 days. Imaging with colposcopy and OCT was performed prior to product use, within 6 h of last dose of product, and 1 week after discontinuation of product. Colposcopy was graded based on vascular and epithelial disruption and erythema. OCT images were evaluated both by quantitative measurement of vaginal epithelial thickness and by use of a qualitative scoring system developed in the macaque to determine epithelial disruption and injury. Mixed model and significance of p=0.05 were used in data analysis.

Results

Baseline colposcopy and OCT findings were similar between treatment groups. After treatment, there were no significant colposcopic differences in vascular or epithelial disruption between treatment groups, with only an increase in erythema noted after treatment in the N-9 group (p=0.01). OCT detected differences between groups in OCT scores (p<0.0001) and epithelial thickness, (p=0.008), both indicators of epithelial injury, after treatment with N-9. One week after discontinuation of treatment, OCT scores were similar between treatment groups (p=0.66) but epithelial thickness in the N-9 group was significantly thicker (p=0.0003).

Conclusions

OCT can be used in a clinical setting to detect epithelial injury and is more sensitive than colposcopy to detect microscopic epithelial disruption. In addition, it gives information about the thickness of the epithelium, a measure previously available only through invasive biopsy. Vaginal epithelium thinned after treatment in the N-9 group, but then thickened after discontinuation of N-9, possibly due to post injury epithelial proliferation. OCT should be considered for inclusion in clinical trials for the detection of product-related toxicity to the vaginal and cervical epithelium.
P3-S7.18 Aetiological agents of infective vaginal discharge among women attending a STD clinic in Kumasi, Ghana
Authors: Agyarko-Poku T
Published: July 2011

Objective

To determine the prevalence of aetiological agents in Infective Vaginal Discharge among women attending a sexually transmitted infection (STI) clinic in Kumasi, Ghana. Methodology Three hundred (300) women consisting of 150 sex workers (SW) and 150 non-sex workers (NSW), attending the Suntreso STI clinic in Kumasi, with complaint of vaginal discharge were recruited for the study. Specimens for wet mount, pH determination, whiff test, Gram's stains, culture and polymerase chain reaction (PCR) were collected from the vagina and the cervix for microbial identification. The HIV status of the women was also determined by Western Blot Assay. Details regarding socio demographic characteristics of the women, symptoms and signs as well as sexual behaviour were recorded. Associations of these factors with each of the aetiological agent was recorded and adjusted for other risk factors.

Result

Bacterial vaginosis was the most common cause of infective vaginal discharge (111/300: 37.0%, p =0.000, SW-61/150{40.7%}; NSW–58/150{38.0%} in the women, followed byCandida spp.(99/300: 32.7%, p=0.000, SW-41/150{27.3%}; NSW-58/150{38.0%}),Trichomonas vaginalis(20/300: 6.7%, p=0.000) SW-12/150{8.0%;NSW–8/150{5.3%}),Chlamydia trachomatis(9/300: 3.0%, p=0.001 SW–6/150{4.0%}; NSW-3/150{150{2.0%}),Neisseria gonorrhoeae(6/300: 2.0%, p=0.014, SW–4/150{2.7%}; NSW–2/150{1.3%}) andMycoplasma genitalium(10/300, 3.3%, p=0.000, SW-7/150{2.0%}). 11.3% (34/300, 30 sex workers, four non-sex workers) of the women were HIV antibody positive. All of the aetiological agents exceptChlamydia trachomatis(p=0.705) were associated with HIV infection. There was no difference in the types of aetiological agent found in sex workers (SW) and non-sex workers (NSW). Prevalence of all the aetiological agents was higher among sex workers except forCandida spp.(27.3%, 41/150 compared with non-sex workers (38.0%, 57/150). Younger age (15–29 years) was found to be the strongest predictor of infection.

Conclusion

Agents of Bacteria vaginosis,Candida spp.andTrachomonas vaginaliscontinue to be the most predominant aetiological agents responsible for infective vaginal discharge among women in Kumasi, Ghana, while prevalence ofNeisseria gonorrhoeaeandChlamydia trachomatisdecline. Improving the detection and management of these organisms has significant public health implication for STI and possibly HIV control.
P4-S1.02 Coupling of electrochemical detection with PCR amplification for sensitive detection ofNeisseria gonorrhoeae
Authors: Sood S , Verma R , Singh R , Sumana G , Sharma V K , Samataray J C , Pandey R M , Malhotra B D
Published: July 2011

Background

Despite the recent development of different detection methods, better diagnostic tools are required for quick and reliable detection of pathogens. Use of biosensors for detection of pathogens is gradually gaining momentum. However, PCR amplification of DNA target is still necessary for application on biosensor for accurate detection of the pathogen. An in-house PCR using self-designed primers targeting the opa gene (GenBank accession no. PUID 9716120 SNUM 2706 Ng_opa) was performed. The generated amplicons were used to evaluate the DNA biosensor utilising a 19-mer oligonucleotide sequence (GenBank PUID SNUM: 9716119 2705 Ng_opa) as probe.

Methods

In-house PCR was standardised and an amplified product (amplicons) of 188 bps were obtained for positive samples. Fabrication of bioelectrode was performed by immobilising the activated probe onto pre-treated screen-printed gold electrodes. The fabricated nucleic acid functionalised gold electrode was characterised using, SEM, CV, DPV techniques. The presence of target DNA was detected electrochemically by monitoring the redox peak of methylene blue indicator. Standardisation of working conditions was done using complementary, non-complementary, one base mismatch DNA, and amplicons from standard strain of N. gonorrhoeae (ATCC 49226) & 16 clinical isolates. In addition, DPV measurements of hybridised bioelectrode with amplicons of 26 clinical samples of which 10 were culture positive, was done. A cut-off value for positives was determined by using the software STATA (version 9).

Results

The analytical sensitivity of PCR was 10–17 M of DNA and the bioelectrode could detect up to 1.0×10–20 M of the DNA amplicons. An 11.49% decrease in signal intensity was taken as the cut-off. Samples giving an equivalent or more decrease than this value were considered as positives.

Conclusions

The coupling of electrochemical detection with PCR amplification showed the advantage of higher sensitivity and increased specificity for detection ofN gonorrhoeae. This may prove to be particularly valuable for the identification of asymptomatic infections and could greatly improve gonorrhoea control.
P4-S1.03 Demonstrating performance of a low-cost, ultra-rapid PCR device with true point-of-care applications
Authors: Pearce D , Shenton D , Brake S , Larry A , Green M , Gaydos C
Published: July 2011

Background

We are developing a highly sensitive, ultra-rapid, multiplex PCR method with fully-integrated DNA preparation and ambient-stable reagents. The assay was used in conjunction with a novel electrochemical detection method to demonstrate low copy number amplification and detection in <20 min, as a point-of-care (POC) diagnostic test forChlamydia trachomatis(CT).

Methods

The method employs custom PCR cards, utilising a thin-film laminate construction to achieve rapid heat transfer, in conjunction with an ultra-rapid thermocycler. All reagents necessary to perform the extraction, amplification and detection are deposited into the cards and air dried at the point of manufacture. Novel, ambient-stable reagent formulations with an 18 month shelf life have been developed. A sample is added to the card and DNA extracted from the sample. The resulting eluate reconstitutes dried PCR reagents and a 40-cycle multiplex PCR is performed using rapid thermocycling. Amplified target is detected using electrochemically-labelled target-specific probes and a double-stranded DNA-specific exonuclease to release the electrochemical label. Released label is read by applying a voltage to a screen printed carbon electrode and at a known oxidation potential the label is oxidised producing a measurable current. The unique rapid performance of this device has been demonstrated in terms of analytical sensitivity and reagent stability under ambient storage conditions. Multiplex capability is demonstrated in this test with the presence of internal control (IC) DNA.

Results

Analytical sensitivity of the device was evaluated by testing dilutions of CT in the presence of IC DNA. The results show CT detection down to 50 copies when co-extracted, amplified in duplex and detected electrochemically with the IC DNA (see graph). Tests on the reagents dried into the device showed stability for 18 months when stored at ambient temperature (20–25°C). Reagent performance after 18 months' storage was shown to be equivalent to performance at time zero see Abstract P4-S1.03 figure 1.

Abstract P4-S1.03 Figure 1

Conclusions

The results show that this device could be used to perform ultra-rapid multiplex PCR with no user intervention after sample addition, allowing minimally-trained staff to carry out the assay in <20 min, meeting the needs for a “true” POC device. Ambient stability of the reagents negates the requirement for any specialised storage conditions.
P4-S1.07 Chlamydia trachomatisDNA stability independent of preservation temperature, type of medium en storage duration
Authors: van Dommelen L , van Tiel F H , Dukers N H , Wolffs P F , Herngreen S B , Hoebe C J P A
Published: July 2011

Background

In almost all diagnostic studies, stored samples are used. This is also the case inChlamydia trachomatisresearch. The performance of diagnostic assays, such as nucleic acid amplification tests, is dependent on the preservation ofC trachomatisDNA. However, the influence of different storage conditions onC trachomatisDNA has not been thoroughly explored yet. Therefore, we have studied the impact of temperature, type of medium and duration of storage on the preservation ofC trachomatisDNA.

Methods

Phosphate buffered saline (PBS), 2-sucrose-phosphate medium (2-SP), COBAS Amplicor medium and urine samples were spiked withC trachomatisserovar D elementary bodies and stored at room temperature, 4°C, −20°C and −80°C. DNA was isolated using the Qiagen DNA mini kit (Qiagen GmbH, Hilden, Germany). Samples were tested in triplicate with a TaqMan PCR at day 0, 1, 7, 14 and 30. Furthermore, clinicalC trachomatispositive urine samples were collected and pooled, and stored and tested in triplicate, at the same temperatures and time intervals. This same procedure was executed with pooledC trachomatispositive swab samples.

Results

C trachomatiswas detected in all samples, irrespective of the different storage temperature conditions, different types of medium and the different durations of storage. Regarding storage time, the cycle threshold (ct) overall did not increase, in fact it tended to decrease within 30 days. If there was an increase in ct over time, this occurred in <10% of the experiments, it did not outreach three (corresponding to a decline in load of 1 log). This mainly occurred in the spiked COBAS samples which were frozen.

Conclusions

All samples tested positive forC trachomatisin our experiments, with on average no decrease in cycle threshold over time. We can therefore conclude that PBS, 2-SP, COBAS Amplicor medium and urine are all excellent media to preserveC trachomatisDNA for a longer period of time, independent of storage temperature.