In 1996, a RCOG Study Group reporting on the prevention of pelvic infection highlighted the considerable role <i>C. trachomatis </i>played in female reproductive morbidity and the potential advantages of DNA based assays.  A national screening programme was suggested, as Sweden and the USA had demonstrated that screening women could decrease prevalence and pelvic inflammatory disease rates. In the UK, out with genito-urinary medicine clinics, awareness of the infection and screening was virtually non-existent.  Women attending obstetric and gynaecology-affiliated clinics were at increased risk of ascending infection compared to the general public and ideally placed for opportunistic screening.  However, patients were <i>TESTED </i>only if symptomatic, by specimens taken from the endocervix for culture or antigen detection assay.  It was from this background that the studies commenced.  The thesis comprises of: -  A questionnaire survey assessing sexually active women’s knowledge of <i>C. trachomatis </i>infection and perceived acceptability of different methods and settings for screening.  Women attending induced abortion and family planning clinics in Aberdeen and Leeds were recruited. -  A prevalence study, aiming to identify who should undergo screening.  Sexually active women attending six different clinical settings in Aberdeen’s Obstetrics & Gynaecology department were screened for <i>Chlamydia.</i> -  A study assessing test performance and acceptability of four different screening approaches (enzyme immunoassay of endocervical specimens and ligase chain reaction assay of endocervical, clinician-collected vulva!, and urine specimens) to opportunistically screen pregnant and non-pregnant women, under 25<i> </i>years of age. -  A study evaluating patient-collected vulval swabs, as an alternative to non-invasive screening by urine.  Women under 25 years of age attending a family planning clinic were opportunistically screened and test performance and acceptability evaluated. -  A study determining whether the measurement of chlamydial IgG antibodies alone or in combination with medical history and/or transvaginal ultrasound can predict tubal infertility in subfertile women.

Diss. (sammanfattning) Uppsala : Uppsala universitet, 2006. / Härtill 5 uppsatser.

<p>The objectives of this case-control study, investigating 51 in-patient women with acute pelvic inflammatory disease (PID) and 50 healthy women attending for routine gynecological check-up, were to investigate the background and reproductive history of women, who are considered at low risk of sexually transmitted infection presenting with PID, to examine whether intrauterine device (IUD) use <i>per se</i> and long use affects the microbiology and clinical course of the disease, to identify risk factors for PID and to examine whether IUD use is an independent risk factor for PID.</p><p>The most striking difference regarding the background and reproductive history between women with PID and healthy women over age 25 were related to socio-demographic factors and not to common risk factors for PID.</p><p>There was little difference between healthy women and women with current PID with regard to single microbes. The finding of combinations of several anaerobic or aerobic/anaerobic microbes appeared to be associated with PID, particularly in women over 35. The pathogenesis of non-sexually transmitted PID appears to be associated with a synergistic effect between several pathogens, possibly facilitated by the presence of an IUD.</p><p>IUD use <i>per se</i> was associated with an increased risk of PID in women 35 and older. There was an association between IUD use and complicated PID in women over 35, which was possibly influenced by long-term IUD use. Age over 35 and IUD use, independently of each other, were associated with an increased risk of failed conservative treatment, necessitating surgery in patients with PID.</p><p>An observational study showed that Latvian obstetrician-gynecologists participate actively in contraceptive counseling and are very experienced with regard to IUD use. Physicians’ attitudes and perceptions towards IUD are generally positive and their clinical considerations are in good agreement with that of doctors in other countries. Antibiotics are widely used around IUD insertion by doctors, possibly driven by a liberal attitude towards IUD use in women with a potential risk of STI. The study could identify some possible gaps in the theoretical knowledge about the IUD and other methods.</p>

The objectives of this case-control study, investigating 51 in-patient women with acute pelvic inflammatory disease (PID) and 50 healthy women attending for routine gynecological check-up, were to investigate the background and reproductive history of women, who are considered at low risk of sexually transmitted infection presenting with PID, to examine whether intrauterine device (IUD) use per se and long use affects the microbiology and clinical course of the disease, to identify risk factors for PID and to examine whether IUD use is an independent risk factor for PID. The most striking difference regarding the background and reproductive history between women with PID and healthy women over age 25 were related to socio-demographic factors and not to common risk factors for PID. There was little difference between healthy women and women with current PID with regard to single microbes. The finding of combinations of several anaerobic or aerobic/anaerobic microbes appeared to be associated with PID, particularly in women over 35. The pathogenesis of non-sexually transmitted PID appears to be associated with a synergistic effect between several pathogens, possibly facilitated by the presence of an IUD. IUD use per se was associated with an increased risk of PID in women 35 and older. There was an association between IUD use and complicated PID in women over 35, which was possibly influenced by long-term IUD use. Age over 35 and IUD use, independently of each other, were associated with an increased risk of failed conservative treatment, necessitating surgery in patients with PID. An observational study showed that Latvian obstetrician-gynecologists participate actively in contraceptive counseling and are very experienced with regard to IUD use. Physicians’ attitudes and perceptions towards IUD are generally positive and their clinical considerations are in good agreement with that of doctors in other countries. Antibiotics are widely used around IUD insertion by doctors, possibly driven by a liberal attitude towards IUD use in women with a potential risk of STI. The study could identify some possible gaps in the theoretical knowledge about the IUD and other methods.

The asymptomatic nature of chlamydial infection renders the differential diagnosis of acute and chronic infection difficult. An untreated Chlamydia trachomatis infection can become chronic, result in disease sequelae such as salpingitis and pelvic inflammatory disease (PID), and ultimately culminate in tubal occlusion and infertility. Diagnostic tests for C. trachomatis such as nucleic acid amplification testing (PCR), antigen detection and serological methods have variable performance capabilities with respect to sensitivity, specificity and stage of infection. The use of PCR as a diagnostic tool is somewhat limited, as specimen collection is routinely sampled from the lower genital tract; hence, infections in the fallopian tube where inflammatory damage is most significant, escape detection. Furthermore, PCR can only detect selected Chlamydia DNA sequences from readily accessible sites of the genital tract, and therefore cannot differentiate between acute and chronic infection. Other serological assays aim to discriminate the various stages of C. trachomatis infection through identification of key antigens. The efficacy of these assays however is impeded due to cross-reactivity between chlamydial species and the subsequent antibody response against the target antigen is not restricted to patients with a specific stage of infection. To identify antibody responses capable of differentiating various states of chlamydial infection, samples were collected from both men and women given the variability of immune responses between the two genders. Samples were assigned to a patient group according to infection status and then probed against protein extracts of HEp-2 cells infected with C. trachomatis serovar L2 and HEp-2 cells pre-treated with IFN-γ and infected with C. trachomatis serovar L2. (persistence cell culture) Serological analysis revealed the presence of five antigens (denoted bands A, B, C, D and M) which were shown to be differential between patient groups. Identification of bands B and C by N-terminal sequencing provided two possible candidates for each antigen, ie. CT727 and CT396 (band B) and CT157 and CT423 (band C). In contrast, band M which was unique to males was a PmpB (probable outer membrane protein B) fragment. The four target antigens (CT157, CT423, CT727 and CT396) were expressed as recombinant proteins using autoinduction media and were subsequently probed by both male and female sera to evaluate their diagnostic potential. Results showed that two chlamydial antigenic targets (CT157 and CT727) have the potential to discriminate between acute and chronic C. trachomatis infection. However, since only a small number of samples (n = 3) were used for this aspect of the study, the findings should simply be viewed as preliminary. In females, sensitivity and specificity values were derived using various combinations of the four target antigens into a panel format for the purpose of detecting chronic C. trachomatis infections. The preferred format was B + C with a sensitivity and specificity of 80% and 84% respectively. Using the IFN-γ-mediated persistence model, only two of the five antigenic targets were shown to be differentially expressed. PmpB in males and CT157 (the most likely band C candidate) in females were shown to be up-regulated to varying degrees in samples across the patient groups. We also demonstrated that no other chlamydial antigens are up-regulated during a persistent C. trachomatis infection. In conclusion, although combinations of bands A, B, C, D and M differentiate between male and female patient groups under normal chlamydial growth conditions, during IFN-γ-induced persistence, only bands C (CT157) and M (CT413 - PmpB) are up-regulated thus suggesting a potential role in chronic C. trachomatis infection.

Philosophiae Doctor - PhD / Sexually transmitted infections (STIs) are a problem for both developed and developing countries. Sub-Saharan Africa has the highest rates in the 15-49 years old group. The discovery that these infections playa vital role in the transmission of HIV raised their profile and made their control one of the central strategies of stopping the HIV/AIDS epidemic. In response to the challenge of improving the quality of care for people infected with STIs in the public health sector, the South African Ministry of Health adopted the syndromic management approach, recommended by the World Health Organisation as suitable for resource-poor settings, for use in primary health care clinics. In addition to providing guidelines on clinical management of STIs, the syndromic approach requires health providers to counsel and educate patients about STIs, encourage patients to complete treatment even if symptoms abate, promote condom use and the treatment of all sexual partners. While the management guidelines are clear and detailed around the diagnostic and medication issues, the processes of education and counseling are not as clearly outlined. Furthermore, although the syndromic approach is a viable way of providing good quality care to larger sections of the population than could be serviced through dedicated STI clinics, it requires health providers working in primary health care clinics, most of whom are professional nurses, to perform some tasks for which they may not be adequately trained. This study evaluated the quality of care for persons infected with ST!s by examining the extent to which the syndromic approach was being implemented in primary health care clinics. Interviews, using semi-structured interview schedules, were conducted with ST! patients and health providers in twenty-four clinics located in four provinces. In depth qualitative interviews were also conducted with a sub-sample of the patients. For further triangulation the methods of participant observation, through the use of simulated patients, and focus group discussions with various community groups were used. The findings of the study indicate that although primary health care clinics in South Africa are well-resourced, the management of patients with ST!s is inadequate. Adherence to the various aspects of syndromic management was poor. Similar to other studies in South Africa, the attitudes of health providers towards patients with ST!s were found to be problematic, a finding that has implications for health-seeking behaviours. The thesis argues that a large part of the problem is related to the multiple roles that nurses have to play in primary health care settings, as well as the content and methodology of the training of nurses who manage ST! patients. It further argues for the constitution of the basic health team at primary health clinics to be multi-disciplinary, and for a multi-disciplinary input in the training of health providers.

Objective: The clinical significance of the potential for persistent human chlamydial infections in vivo is being actively reassessed because of the increased frequency of recurrent infection with the same serovar despite compliance with an effective antibiotic regimen. The ability to extend the length of time of in vitro cultivation of polarized human endometrial epithelial cells (HEC-1B) provided the opportunity to establish a model system to determine if a persistent form of Chlamydia trachomatis had the same susceptibility as the actively growing form to a cidal concentration of azithromycin. Methods: Polarized HEC-1B cells cultivated on extracellular matrix were infected with C. trachomatis serovar E and exposed to penicillin at 24 h post-infection (hpi) to induce a persistent infection characterized by slowly metabolizing but non-dividing, ultrastructurally aberrant reticulate bodies within the chlamydial inclusion; at 48hpi, infected cultures were exposed to a bactericidal concentration of azithromycin for 72 h. Results: Persistent chlamydiae were phenotypically resistant to azithromycin; the number of chlamydial inclusions on subpassage of progeny from persistent chlamydiae following removal of penicillin and recovery was essentially the same as from progeny from persistent chlamydiae following removal of penicillin and azithromycin and recovery. Neutrophils were attracted in vitro to persistently infected HEC-1B cells that had been exposed to penicillin and azithromycin. Conclusions: Thus, this study provides evidence at the cellular microbiology level in vitro for mechanisms that could exist in vivo to create sustained, but perhaps clinically inapparent inflammation, which might eventually lead to conditions such as silent pelvic inflammatory disease.

This study is about evaluating the effectiveness of training in syndromic management ofSTDs. The purpose of this study was to fmd out to what extent the STD training programme had been implemented. A stratified sample of twenty-two primary health care clinics in the Port Shepstone region was used. Twenty-two professional nurses rendering STD management in the clinics were observed. Data-gathering was done through a clinic inventory obtained by interviewing the sister-incharge, observation of professional nurses providing treatment to STD patients, interviews with professional nurses to assess their knowledge of the syndromic approach, and exit interviews with patients treated at the clinic. The fmdings suggest that certain areas in the STD training need to be emphasized, such as vaginal speculum and bi-manual examinations. It is recommended that certain negative aspects which hinder effective STD management, like staff shortages and lack of equipment be given attention. / Health Studies / M.A. (Health Studies)

This study is about evaluating the effectiveness of training in syndromic management ofSTDs. The purpose of this study was to fmd out to what extent the STD training programme had been implemented. A stratified sample of twenty-two primary health care clinics in the Port Shepstone region was used. Twenty-two professional nurses rendering STD management in the clinics were observed. Data-gathering was done through a clinic inventory obtained by interviewing the sister-incharge, observation of professional nurses providing treatment to STD patients, interviews with professional nurses to assess their knowledge of the syndromic approach, and exit interviews with patients treated at the clinic. The fmdings suggest that certain areas in the STD training need to be emphasized, such as vaginal speculum and bi-manual examinations. It is recommended that certain negative aspects which hinder effective STD management, like staff shortages and lack of equipment be given attention. / Health Studies / M.A. (Health Studies)

Introdução: Chlamydia trachomatis (CT) é causa da infecção sexualmente transmitida de origem bacteriana mais comum. Na mulher, a infecção genital pela CT pode causar cervicite, uretrite, endometrite, salpingite. Infecções persistentes ou recorrentes provavelmente representam um importante fator de risco para o desenvolvimento de sequelas associadas, como dor pélvica crônica, gravidez ectópica e infertilidade por fator tubário. A lectina ligadora de manose (MBL), componente doo sistema imune inato, tem importante papel na defesa antimicrobiana, reconhecendo vírus, fungos e patógenos bacterianos. O gene que codifica para a MBL é polimórfico, e a substituição de um único nucleotídeo resulta na produção de uma proteína instável, que é rapidamente degradada. Objetivo: Avaliar se mulheres brasileiras portadoras de um polimorfismo do gene da MBL apresentam diferentes susceptibilidades para a ocorrência de obstrução tubária, na presença ou ausência de uma infecção prévia por Chlamydia trachomatis. Métodos: Em estudo caso-controle, foram avaliadas 75 pacientes com obstrução tubária e 75 pacientes com tubas pérvias, atendidas na Divisão de Ginecologia do Hospital das Clínicas da Universidade de São Paulo (HCFMUSP). Anticorpos IgG anti-CT foram mensurados através de um ensaio de imunoabsorção enzimática para investigar uma infecção prévia pela CT. Para o polimorfismo, realizou-se coleta de células bucais e o DNA extraído foi analisado através de reação em cadeia da polimerase (PCR), digestão de endonuclease e gel de eletroforese, utilizando pares de primers específicos para a região polimórfica. Todo material foi mantido a -80°C e enviado em gelo seco para a Division of Immunology and Infectious Diseases da Weill Cornell Medicine em Nova York. Associações entre genótipos de MBL ou alelos e permeabilidade tubária foram analisadas pelo teste de qui-quadrado de Pearson com ou sem correção de Yates. Resultados: Não houve diferença na detecção de anticorpos da CT entre os grupos. Mulheres com tubas obstruídas tiveram uma prevalência maior do genótipo AB (36%) versus (16%), resultado estatisticamente significativo (p < 0,01). De maneira semelhante, a distribuição do alelo A e do alelo variante B também apresentaram diferenças significantes entre os grupos (p < 0,01). Conclusão: Os achados sugerem, que embora a exposição à Chlamydia trachomatis tenha sido semelhante em ambos os grupos, a presença do alelo variante B do gene que codifica para a MBL aumenta o risco de desenvolvimento da obstrução tubária, subsequente à infecção pela CT ou outros agentes infecciosos. Nas mulheres brasileiras avaliadas a presença de tal polimorfismo genético aumentou a probabilidade de obstrução tubária em consequência de uma infecção do trato genital / Introduction: Chlamydia trachomatis (CT) is the cause of the most common bacterial sexually transmitted infection. In women, genital CT infection may cause cervicitis, urethritis, endometritis, salpingitis. Persistent or recurrent infections probably represent an important risk factor for the development of associated sequelae, such as chronic pelvic pain, ectopic pregnancy and tubal factor infertility. Mannose-binding lectin (MBL), a component of the innate immune system, has an important role in antimicrobial defense, recognizing viral, bacterial and fungal pathogens. The gene coding for MBL is polymorphic and a single nucleotide substitution results in production of an unstable protein, that is rapidly degraded. Objective: To evaluate whether Brazilian women with a polymorphism in the MBL gene present different susceptibilities to the occurrence of fallopian tube damage, in the presence or absence of a previous infection by CT. Method: In a case-control study, 75 patients with tubal obstruction and 75 patients with patent tubes were studied, all seen at the Gynecology Division of the Hospital das Clínicas of the University of São Paulo (HCFMUSP). IgG anti-CT antibodies were measured by enzyme-linked immunoassay to investigate a previous CT infection. For the polymorphism analysis, buccal cells were collected and the extracted DNA was analyzed by polymerase chain reaction (PCR), endonuclease digestion and gel electrophoresis using primer pairs specific for the polymorphic region. All material was maintained at -80 ° C and sent on dry ice to the Division of Immunology and Infectious Diseases at Weill Cornell Medicine in New York. Associations between MBL genotypes or alleles and tubal permeability were analyzed by the Pearson chi-square test with or without Yates correction. Results: There was no difference in CT antibody detection between the two groups. Women with obstructed tubes had a higher prevalence of being positive for the heterogenous genotype AB (36%) versus (16%) (p < 0.01). Similarly, the distribution of the normal A allele and variant B allele were also significant different between the two groups (p < 0,01). Conclusion: The findings suggest that while exposure to CT was similar in both groups of women the presence of the variant MBL B allele increases the risk for development of tubal obstruction, subsequent to a CT or other infection. In the Brazilian women evaluated possession of this genetic polymorphism increased the likelihood that blocked fallopian tubes will be a consequence of a genital tract infection