Diss. (sammanfattning) Stockholm : Karol. inst., 2006. / Härtill 5 uppsatser.

Neisseria gonorrhoeae (NG) har successivt utvecklat resistens mot många antimikrobiella medel och betraktas som ett av de tre reella hoten bland antibiotikaresistenta bakterier. Ciprofloxacin är ett bredspektrum-antibiotikum tillhörande gruppen kinoloner som, förutom att behandla urinvägsinfektioner, används mot NG och infektioner i mage och tarm. Dock har det rapporterats att ca 30 % av NG-isolat som samlats in genom gonokock-isolatövervakningsprojekt (GISP) under 2017 var resistenta mot ciprofloxacin. På molekylnivå är resistens mot ciprofloxacin starkt associerad med en enda mutation i kodon 91 i gyras-genen (gyrA). Detta projekt har undersökt om det går att använda molekylärbiologiska metoder för att detektera NG-isolat med gyrA mutationen. Analysen gjordes med två olika PCR-system, ”7500 Fast Real-Time PCR System” från Applied Biosystems (ABI) och Panther Fusion från Hologic. Proberna som användes designades för påvisning av vildtyp gyrA (ciprofloxacin-känslig) och mutant gyrA (ciprofloxacin-resistent) hos NG. I projektet analyserades 50 NG-positiva prov (analyserade med screeningtest APTIMA COMBO2 från Hologic), från 43 patienter som provtagits under januari-februari 2020 i Region Skåne. Några patienter testades flera gånger vid olika tillfällen. NG-odling hade utförts parallellt från motsvarande tagna prov från patienterna. ABI-metoden påvisade genen hos 90 % (45/50) av NG-positiva prover (APTIMA COMBO2) medan endast 24 av de 49 proven (49 %) kunde odlas med traditionell metodik för att därefter resistensbestämmas. Av de 45 prov där gyras-genen kunde detekteras med ABI-metoden, uppvisade 28 (62 %) av proven en muterad gen och därmed en potentiell resistens för ciprofloxacin. Panther Fusion-metoden påvisade genen hos 80 % (40/50) av NG-positiva prover (APTIMA COMBO2), och såsom tidigare nämnts, kunde endast 24 av de 49 proven (49 %) odlas med traditionell metodik för att därefter resistensbestämmas. Av de 40 prov där gyras-genen kunde detekteras med Panther Fusion-metoden, uppvisade 26 av proven (65 %) en muterad gen och därmed en potentiell resistens för ciprofloxacin. En jämförelse mellan resultaten från PCR-metoderna och odlingarna visar att av de 24 odlingarna som kunde resistensbestämmas fick ABI-metoden resultat för 23 och Panther Fusion för 22. PCR-metodernas resultat överensstämde perfekt med resultaten från odling med samma 8 känsliga och 15 respektive 14 resistenta NG-isolat som odling. De båda PCR-metoderna och traditionell odling uppvisade jämförbara resultat. Av de 24 prov som kunde odlas och därmed resistensbestämmas, detekterades med ABI-metoden gyras-genen i 23 av dessa prov och i 22 av proven med Panther Fusion-metoden. Resistens mot ciprofloxacin uppvisades genom odling i 16 av de 24 odlingsbara prov, och av dessa 24 odlingsbara prov uppvisade ABI-metoden en muterad gen i 15 av proven och Panther Fusion-metoden en muterad gen i 14 av proven. Traditionell odling kunde bara genomföras på 24 av proven och PCR-metoderna identifierade signifikant fler prov innehållande vildtyp eller muterad gyras-gen, 45 respektive 40 prov. Projektet visade tydligt att PCR-metoderna kan identifiera fler prov än genom traditionell odling och kan därmed upptäcka fler prov med förväntad ciprofloxacin-resistens än vad som kan bestämmas genom traditionell odling. / Neisseria gonorrhoeae (NG) has been developing a resistance towards several different antibiotics and is viewed as one of the three real threats among resistant bacteria. Ciprofloxacin is a broad-spectrum-antibiotic belonging to the group quinolone antibiotics which, in addition to being used to treat urinal infections, is used to treat NG and infections in the stomach and intestines. However, it has been reported that 30 % of NG-isolates that have been gathered through the Gonococcal Isolate Surveillance Project (GISP) throughout 2017 were resistant to ciprofloxacin. On a molecular level, resistance to ciprofloxacin is strongly associated with a single mutation in kodon 91 in the gyras-gene (gyrA). This project sought to examine if it is possible to use methods from molecular biology to detect which NG that have the gyrA-mutation. The test was done using two different PCR-systems, ”7500 Fast Real-Time PCR System” from Applied Biosystems (ABI) and Panther Fusion from Hologic. The probes used were designed to show wild type gyrA (ciprofloxacin sensitive), and mutated gyrA (ciprofloxacin resistant) in NG. In this project 50 NG-positive samples (analysed with screentest APTIMA COMBO2 from Hologic), from 43 patients that had been tested during January-February 2020 in Region Skåne, were analysed. Some patients were tested several times, within the time period. NG-cultivation had been done in parallel from corresponding samples taken from the patients. The ABI-method showed the gene in 90 % (45/50) of NG-positive samples (APTIMA COMBO2) while only 24 of the 49 samples (49 %) could be cultivated by traditional methodology, and then tested for resistance. Of the 45 samples where the gyras-gene could be detected with the ABI-method, 28 samples (62 %) exhibited a mutated gene and thus a potential resistance to ciprofloxacin. The Panther fusion-method showed the gene in 80 % (40/50) of NG-positive samples (APTIMA COMBO2), and as mentioned earlier, only 24 of the 49 samples (49 %) could be cultivated by traditional methodology to then be tested for resistance. Of the 40 samples where the gyras-gene could be detected with the Panther Fusion-method, 26 samples (65 %) exhibited a mutated gene and thus a potential resistance to ciprofloxacin. The two PCR-methods and traditional cultivation exhibited comparable results. Of the 24 samples that could be cultivated and thus tested for resistance, the ABI-method detected the gyras-gene in 23 of these samples and the Panther Fusion-method detected the gene in 22 of the samples. Cultivation exhibited resistance to ciprofloxacin in 16 of the 24 samples that could be cultivated, and of these 24 cultivatable samples the ABI method exhibited a mutated gene in 15 of the samples and the Panther Fusion-method exhibited a mutated gene in 14 of the samples. Traditional cultivation could only be done on 24 of the samples and the PCR-methods could identify significantly more samples containing either wild type or mutated gyras-gene, 45 and 40 samples, respectively. The project clearly showed that more samples can be identified with the PCR-methods than through traditional cultivation, and thereby discover more samples with expected ciprofloxacin-resistance, than can be determined through traditional cultivation.

Neisseria gonorrhoeae verursacht die sexuell übertragbare Krankheit Gonorrhoe und ist ein Typ-IV-Pili (Tfp) exprimierendes Bakterium, das den Urogenitaltrakt besiedelt. Frühe Infektionsstadien piliierter N. gonorrhoeae (P+GC) sind durch die Tfp-vermittelte Adhärenz an Wirtszellen gekennzeichnet, dann erfolgt die Bildung von Mikrokolonien auf Wirtszellepithelien. Hier wird gezeigt, dass die Wirtszellen an der effizienten Bildung der extrazellulären Mikrokolonien beteiligt sind. P+GC die fixierte Wirtszellen infizieren weisen eine verzögerte Mikrokoloniebildung gegenüber einer Infektion lebender Wirtszellen auf. Kortikales Aktin wird zusammen mit Signalproteinen innerhalb der Wirtszellen zu den adhärierten Bakterien rekrutiert, darunter das Hauptstrukturprotein von Caveolae-Membrandomänen, Caveolin-1 (Cav1). Eine Reduzierung der Expression von Cav1 führt zu einer verstärkten Aufnahme von P+GC in die Wirtszellen, wohingegen die Expression von Cav1 in Cav1-negativen Zellen eine Internalisierung verhindert. Internalisierte Bakterien weisen dabei geringere Überlebensraten auf je länger sie in den Wirtszellen verbleiben. Die Rekrutierung von Cav1 ist eine unmittelbare und kontinuierliche zelluläre Antwort auf eine Infektion mit P+GC, welche die Phosphorylierung von Cav1 an Tyrosin 14 bedingt. Zusätzlich erforderte die Cav1-vermittelte Blockierung der Internalisierung der Bakterien und die Verankerung von Cav1 mit dem Zytoskelett eine Tyrosinphosphorylierung von Cav1. Eine Analyse möglicher Interaktionspartner von phosphoryliertem Cav1 zeigte eine direkte Interaktion mit Vav2. Sowohl Vav2 als auch sein Substrat, die kleine GTPase RhoA, blockieren die Aufnahme von Bakterien in die in Wirtszellen. Die Aktivierung von RhoA nach P+GC Infektion erfordert die Expression von Cav1, was auf einen Cav1-Vav2-RhoA Signalweg hindeutet. Darüber hinaus wurden in dieser Arbeit sechs neue, eine SH2-Domäne-beinhaltende Interaktionspartner von phosphoryliertem Cav1 identifiziert. / Neisseria gonorrhoeae causes the sexually transmitted disease gonorrhea and colonizes mucosal epithelia of the human urogenital tract. The early stages of infection with piliated N. gonorrhoeae (P+GC) are characterized by Tfp-mediated adherence to host cells, followed by formation of bacterial microcolonies on the surface of host cells. This study provides evidence that host cell participation is required for the efficient formation of extracellular microcolonies during Neisseria infection. P+GC infecting fixed host cells demonstrate altered motility and delayed microcolony formation compared to infecting living host cells. Cortical actin and various signal transducing proteins are recruited to the site of bacterial attachment within host cells, one of them being the major structural protein of plasma membrane caveolae, Caveolin-1 (Cav1). Down-regulation of Cav1 results in increased uptake of P+GC into host cells whereas expression of the protein in Cav1-negative cells blocks bacterial internalization. Host cell entry results in decreased viability of internalized bacteria over time. Cav1 recruitment is demonstrated to be an immediate and continuous cellular response to P+GC infection that involves Cav1 phosphorylation on its tyrosine 14 residue. Prevention of bacterial uptake mediated by Cav1 as well as tight association of Cav1 with the cytoskeleton also requires tyrosine phosphorylation. A broad analysis of interaction partners of phosphorylated Cav1 revealed a direct interaction with the Rho-family guanine nucleotide exchange factor Vav2. Both Vav2 and its substrate, the small GTPase RhoA, are involved in preventing bacterial uptake and RhoA activation after P+GC infection requires Cav1 expression, thus providing evidence for a Cav1-Vav2-RhoA signaling cascade. Moreover, six novel SH2-domain containing interaction partners of tyrosine phosphorylated Cav1 have been identified, all of which have been implicated in modulating the cytoskeleton.

Diss. (sammanfattning) Uppsala : Uppsala universitet, 2007. / Härtill 4 uppsatser.

Made available in DSpace on 2016-12-23T13:56:05Z (GMT). No. of bitstreams: 1 Marcelo Joaquim Barbosa.pdf: 493310 bytes, checksum: 1cc616c331737b5497291791ef93fb83 (MD5) Previous issue date: 2010-07-12 / Objetivos: Determinar a prevalência de Neisseria gonorrhoeae (NG) e Chlamydia trachomatis (CT) e identificar fatores demográficos, comportamentais e clínicos correlacionados a essas infecções em homens atendidos em seis clínicas de DST no Brasil. Métodos: Estudo multicêntrico, em corte transversal realizado em homens que procuraram atendimento em clínicas de DST. O estudo incluiu clínicas de DST em seis cidades distribuídas nas cinco macrorregiões do Brasil, 2004/2005. Metodologia: Coletou-se 20 ml do primeiro jato de urina para testar NG e CT por DNA-PCR. Resultados: Um total de 767 (92,9%) homens foi incluído no estudo. A mediana de idade foi de 24 (DIQ21 30) anos. A prevalência de infecção por CT foi 13,1% (IC95% 10,7%-15,5%) e por NG 18,4% (IC95% 15,7%-21,1%). A prevalência de coinfecção foi 4,4% (IC95% 2,95%-5,85%). Os fatores identificados como sendo independentemente associados com a infecção por clamídia no modelo final de regressão logística foram: ser jovem (15-24 anos) [OR=1,4 (IC95% 1,01-1,91)], apresentar corrimento uretral ao exame [OR=4.8 (IC95% 1,52-15,05)], verrugas genitais [OR=3,0 (IC95% 1,49-5,92)] e história prévia de corrimento uretral [OR=2,4 (IC95% 1,11-5,18)]. As variáveis associadas com gonorréia foram: ser jovem (15-24 anos) [OR=1,5 (IC95% 1,09-2,05)], apresentar corrimento uretral ao exame [OR=9,9 (IC95% 5,53-17,79)], verrugas genitais [OR=18,3 (IC95% 8,03-41,60)] e úlcera ao exame clínico [OR=4,9 (IC95% 1,06-22,73)]. Conclusões: Estes resultados mostram implicações importantes na realização de diagnóstico e tratamento precoces para evitar a transmissão, complicações e implementação de medidas de educação em saúde direcionadas aos homens e deve ser estimulado o oferecimento de testes para DST na rotina dos nos serviços. / Objectives: To determine the prevalence of Neisseria gonorrhoeae(NG) and Chlamydia trachomatis (CT) infections and identified demographic, behavioral, and clinical correlates of these infections in men attending six STI clinics in Brazil. Methods: Multicentric, cross-sectional study performed among men attending STI clinics in Brazil. The study included STD clinics in six diverse cities in the five geographic regions of Brazil in 2005. Patients provided 20 ml of first catch urine for testing for NG and CT by DNA-PCR. Results: A total of 767 (92.9%) men were included in the study. The median age was 24 (IQR 21 30) years old. Prevalence of Chlamydia infection was 13.1% (95%CI 10.7%-15.5%) and gonorrhea was 18.4% (95%CI 15.7%-21.1%). Coinfection prevalence was 4.4% (95%CI 2.95%-5.85%) in men who sought assistance in STI clinics. Factors identified as being independently associated with Chlamydia trachomatis in the final multiple logistic model were being younger (15-24) [OR=1.4 (95%CI 1.01-1.91)], present urethral discharge [OR=4.8 (95%CI 1.52-15.05)], genital warts [OR=3.0 (95%CI 1.49-5.92)] and previous history of urethral discharge [OR=2.4 (95%CI 1.11-5.18)]. Variables associated with gonorrhea were being younger (15 to 24) [OR=1.5 (95%CI 1.09-2.05)], presence of urethral discharge [OR=9.9 (95%CI 5.53-17.79)], genital warts [OR=18.3 (95%CI 8.03-41.60)] and ulcer in clinical examination [OR=4.9 (95%CI 1.06-22.73)]. Conclusions: These findings have important implications for implementing education and prevention efforts directed toward men at risk of HIV/STD. A venue-based approach to offer routinely testing to men in STD clinic should be stimulated.

Made available in DSpace on 2016-12-23T13:56:06Z (GMT). No. of bitstreams: 1 Dissertacao de Marcelo Joaquim Barbosa.pdf: 493316 bytes, checksum: 05c09f00c2dba0603712d1b6f468b750 (MD5) Previous issue date: 2010-07-12 / Objetivos: Determinar a prevalência de Neisseria gonorrhoeae (NG) e Chlamydia trachomatis (CT) e identificar fatores demográficos, comportamentais e clínicos correlacionados a essas infecções em homens atendidos em seis clínicas de DST no Brasil. Métodos: Estudo multicêntrico, em corte transversal realizado em homens que procuraram atendimento em clínicas de DST. O estudo incluiu clínicas de DST em seis cidades distribuídas nas cinco macrorregiões do Brasil, 2004/2005. Metodologia: Coletou-se 20 ml do primeiro jato de urina para testar NG e CT por DNA-PCR. Resultados: Um total de 767 (92,9%) homens foi incluído no estudo. A mediana de idade foi de 24 (DIQ21 30) anos. A prevalência de infecção por CT foi 13,1% (IC95% 10,7%-15,5%) e por G 18,4% (IC95% 15,7%-21,1%). A prevalência de coinfecção foi 4,4% (IC95% 2,95%-5,85%). Os fatores identificados como sendo independentemente associados com a infecção por clamídia no modelo final de regressão logística foram: ser jovem (15-24 anos) [OR=1,4 (IC95% 1,01-1,91)], apresentar corrimento uretral ao exame [OR=4.8 (IC95% 1,52-15,05)], verrugas genitais [OR=3,0 (IC95% 1,49-5,92)] e história prévia de corrimento uretral [OR=2,4 (IC95% 1,11-5,18)]. As variáveis associadas com gonorréia foram: ser jovem (15-24 anos) [OR=1,5 (IC95% 1,09-2,05)], apresentar corrimento uretral ao exame [OR=9,9 (IC95% 5,53-17,79)], verrugas genitais [OR=18,3 (IC95% 8,03-41,60)] e úlcera ao exame clínico [OR=4,9 (IC95% 1,06-22,73)]. Conclusões: Estes resultados mostram implicações importantes na realização de diagnóstico e tratamento precoces para evitar a transmissão, complicações e implementação de medidas de educação em saúde direcionadas aos homens e deve ser estimulado o oferecimento de testes para DST na rotina dos nos serviços / Objectives: To determine the prevalence of Neisseria gonorrhoeae(NG) and Chlamydia trachomatis (CT) infections and identified demographic, behavioral, and clinical correlates of these infections in men attending six STI clinics in Brazil. Methods: Multicentric, cross-sectional study performed among men attending STI clinics in Brazil. The study included STD clinics in six diverse cities in the five geographic regions of Brazil in 2005. Patients provided 20 ml of first catch urine for testing for NG and CT by DNA-PCR. Results: A total of 767 (92.9%) men were included in the study. The median age was 24 (IQR 21 30) years old. Prevalence of Chlamydia infection was 13.1% (95%CI 10.7%-15.5%) and gonorrhea was 18.4% (95%CI 15.7%-21.1%). Coinfection prevalence was 4.4% (95%CI 2.95%-5.85%) in men who sought assistance in STI clinics. Factors identified as being independently associated with Chlamydia trachomatis in the final multiple logistic model were being younger (15-24) [OR=1.4 (95%CI 1.01-1.91)], present urethral discharge [OR=4.8 (95%CI 1.52-15.05)], genital warts [OR=3.0 (95%CI 1.49-5.92)] and previous history of urethral discharge [OR=2.4 (95%CI 1.11-5.18)]. Variables associated with gonorrhea were being younger (15 to 24) [OR=1.5 (95%CI 1.09-2.05)], presence of urethral discharge [OR=9.9 (95%CI 5.53-17.79)], genital warts [OR= 8.3 (95%CI 8.03-41.60)] and ulcer in clinical examination [OR=4.9 (95%CI 1.06-22.73)]. Conclusions: These findings have important implications for implementing education and prevention efforts directed toward men at risk of HIV/STD. A venue-based approach to offer routinely testing to men in STD clinic should be stimulated

Orientadores: Paulo César Giraldo, Ana Katherine da Silveira Gonçalves / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas / Made available in DSpace on 2018-08-16T20:57:25Z (GMT). No. of bitstreams: 1 Amaral_RoseLuceGomesdo_D.pdf: 1220448 bytes, checksum: d6f8f0907e1226cc41668603ab978683 (MD5) Previous issue date: 2010 / Resumo: O uso de duchas vaginais é comum em diferentes povos e culturas. Entre as mulheres que a praticam, a maioria a faz após a menstruação, antes ou após a relação sexual, com a finalidade de evitar o odor desagradável ou para eliminar corrimentos, refrescar a genitália ou prevenir a gravidez. Não está claro na literatura se as duchas poderiam causar problemas às usuárias. Objetivo: Verificar se o uso habitual de duchas vaginais associa-se à infecção genital por Chlamydia trachomatis/ Neisseria gonorrhoeae e Papilomavírus humano (HPV) em mulheres profissionais do sexo. Sujeitos e Métodos: Estudo de corte transversal avaliou 200 mulheres - 111 profissionais do sexo (PS) e 89 não profissionais do sexo (NPS) - assistidas em uma Unidade Básica de Saúde de Campinas, São Paulo, Brasil. Todas as mulheres foram entrevistadas e examinadas por um único pesquisador. A anamnese abordou os antecedentes demográficos, higiênicos, sexuais e médicos, como idade, cor, paridade, escolaridade, tabagismo, estado civil, uso de duchas vaginais, idade da primeira relação sexual, número de parceiros sexuais, número de coitos por semana, práticas sexuais, uso de preservativo e lubrificante. Amostras de células cervicais foram coletadas para testes de captura híbrida objetivando Chlamydia trachomatis/Neisseria gonorrhoaea e HPV. Na análise estatística usou-se o teste exato de Fisher ou X2 para as variáveis discretas, e Mann-Whitney para as variáveis não-paramétricas. O estudo foi aprovado pelo Comitê de Ética em Pesquisa (no 902/2009) e contou com a colaboração da Secretaria de Saúde do Estado de São Paulo e Laboratório Digene. Resultado: Cerca de 40% das mulheres investigadas praticavam duchas vaginais três ou mais vezes por semana (61,7% das PS e somente 14,6% das NPS). A infecção por CT/NG foi positiva em 10,5% do total dos casos, 17 casos (15,3%) em PS e em quatro casos (4,5%) das NPS (p=0,01), porém não houve diferenças significativas entre usuárias de duchas vaginais (14,81%) e não usuárias (7,6%), (p=ns). As PS não usuárias de duchas vaginais tiveram quase o dobro de infecção CT/NG que as NPS usuárias de duchas vaginais (13,9% vs 7,7%). O HPV foi positivo em 40,5% dos casos, sendo 55,8% da PS e em 21,3% das NPS (p=0,001), contudo DNA-HPV não foi significativamente diferente (p=0,47) entre PS usuárias de duchas vaginais (54,4%) e em não usuárias (58,1%). HPV de alto risco foi positivo em 16,2% e 11,6% (p=ns) e o HPV de baixo risco em 23,5% e 30,2% (p=ns) em usuárias e não usuárias de duchas vaginais respectivamente. Os HPV de alto e baixo riscos foram encontrados simultaneamente em 14,7% e 16,2% das usuárias e não usuárias de duchas vaginais, respectivamente (p=ns). Conclusão: O uso de duchas vaginais não se associou às infecções genitais por CT/NG e HPV de alto ou baixo grau nas mulheres estudadas (profissionais do sexo e não profissionais do sexo) / Abstract: The use of vaginal douching (VD) is widespread around the world, and is more common than is to be expected. The majority of women douche after menses, before or after sexual intercourse to prevent odor, to alleviate vaginal symptoms, or to prevent pregnancy, however it is not clear in the literature if this habit can cause damages for women. Objective: Establish whether high frequency VD favors Chlamydia trachomatis (CT)/Neisseria gonorrhoeae (NG) and Papilomavírus humano (HPV) infection in female sex workers (SW). Subjects and Methods: A clinical cross-sectional study involving 200 women, 111 SW and 89 non-sex workers (NSW) in a Health Center in Brazil. The subjects were submitted to an interview and examined by a single researcher. A questionnaire was filled out with data that included, medical and demographic history (age, race, parity, education, smoking, marital status), hygiene (use of VD, frequency and the solution used) and sexual behaviour (first sexual relation, number of sexual partners, frequency, sexual practices, homosexual relationships, use of condom and lubricant). Cervical samples were collected for CT and NG testing by hybrid capture 2 assay. Statistical analysis used the Fisher's exact test or qui square for discrete variables and Mann-Whitney test for nonparametric variables. The study was approved by Committee the Ethics in Research (no 902/2009) and received the cooperation of São Paulo Secretariat of Health and Digene laboratory. Results: Approximately 40% of women practiced douche three or more times per week (61.7% of SW and only 14.6% of the NSW). Infection with CT / NG was positive in 10.5% of the total cases, 17 cases (15.3%) in SW and in four cases (4.5%) of the NSW (p = 0.01). However, It was in 14.81% of D and but in and 7.6% of ND (p = ns). SW douchers had almost double CT and NG infections than NSW douchers (13.9% vs 7.7%). HPV infection was detected in 40.5% of cases, 55.8% of SW and 21.3% of NSW of (p = 0.001), despite of DNA HPV was not different (p = 0.47) between SW douchers (54.4%) and non-douchers (58.1%). High-risk DNA-HPV was positive in 16.2% and 11.6% (p = ns) and low-risk DNA-HPV in 23.5% and 30.2% (p = ns) in douchers and non-douchers respectively. High and low risk DNA-HPV were found simultaneously in 14.7% and 16.2% of douchers and non-douchers respectively (p = ns). Conclusion: The use of VD is not a causal factor for cervical CT/NG and High-/low risk HPV infection in women studied (SW and NSW) / Doutorado / Tocoginecologia / Doutor em Tocoginecologia

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No. of bitstreams: 2 Dissertação - Fernanda Alves de Brito e Cardoso - 2010.pdf: 3862835 bytes, checksum: bac9d567036b30752cd19dc0042891c0 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Previous issue date: 2010-12-14 / Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq / The introduction of the nucleic acid amplification tests (NAATs) was a major breakthrough in the screening for the sexually transmitted diseases (STD) caused by Chlamydia trachomatis and Neisseria gonorrhoeae because they are highly sensitive and they can be used with noninvasive specimens, such as urine. The use of urine has made it far easier to test asymptomatic individuals and has also made it possible to perform epidemiological studies in places other than clinical settings. Many studies have shown also that vaginal swab can be used for detection of both infections, however, just the NAAT Aptima Combo 2 has been cleared by Food and Drug Administration for this specimen use. In Brazil, the most widely used NAAT for the diagnosis of chlamydia and neisseria is the kit Amplicor CT/NG (Roche) and, up to date, there isn’t any study which evaluates the use of vaginal swabs. Objectives: To evaluate the performance of the kit AMPLICOR CT/NG (Roche) in the diagnosis of C. trachomatis and N. gonorrhoeae using urine, endocervical and vaginal swabs and to analyze the agreement of results between the different biological specimens. Methods: The target population was sexually active adolescents and young women between 15 and 24 years from Inhumas, Goias. Socio-demographic and sexual behavior were obtained through a face-to-face interview. The diagnosis was performed by PCR using the AMPLICOR CT/NG (Roche) assay in urine, vaginal swab (VS) and endocervical swab (ES) specimens. For the performance evaluation were calculated the sensitivity, specificity, positive predictive value and negative predictive value. The kappa coefficient was calculated to assess agreement between the samples. It was considered a true-positive result when at least two of three biological samples from the same patient were positive for chlamydia and/or gonococcus. Results:Among the 428 participants the mean age was 19,4 years. The three biological specimens were collected from 309 adolescents (72.2%). Among these, the prevalence rates were 8.7% (IC95% 5,8-12,4) for C. trachomatis and 2.3% (IC95% 0,9-4,6) for N. gonorrhoeae.For chlamydia the sensitivities observed with the different samples were above 80% and specificities exceeding 97% with positive predictive values (PPV) between 78.8% and 84.6% and negative predictive values (VPNs) >98%. For the gonococcus the sensitivities were 42.8% for urine, 71.4% for ES and 100% for VS with specificities >96% for the three samples. The two types of swab showed low PPVs for gonococcus (≈40%) and urine showed PPV of 100%. VPNs were >98%. The agreement of results between specimens was around 94% for the detection of both infections. However, the values of kappa (κ) coefficient ranged from 0.68 to 0.73 for chlamydia, which means substantial agreement between samples. For gonococcal infection, the agreement was slight or fair with κ coefficients ranging from 0.13 to 0.33. Conclusions:The performances of the specimens and the κ values suggest that the vaginal swab appears to be equivalent to urine and endocervical swab for detection of chlamydia and may be suitable for screening studies. The three samples showed different performance in the detection of gonococcus and did not present good agreement of results, suggesting that they are not equivalent in the diagnosis of this infection with the PCR kit used. / A introdução dos testes de amplificação de ácidos nucléicos (NAATs) foi um grande avanço no rastreamento das doenças sexualmente transmissíveis (DST) causadas por Chlamydia trachomatis e Neisseria gonorrhoeae, pois apresentam alta sensibilidade e permitem a utilização de amostras de coleta não invasiva, como a urina. O uso da urina facilitou o diagnóstico em indivíduos assintomáticos e possibilitou a realização de estudos epidemiológicos em locais fora do ambiente clínico. Vários estudos mostram que o swab vaginal também pode ser utilizado na detecção de ambas as infecções, porém a Food and Drug Administration restringe o seu uso apenas pelo NAAT Aptima Combo 2. No Brasil, o NAAT mais utilizado no diagnóstico de clamídia e neisseria é o kit Amplicor CT/NG (Roche) e, até o momento, nenhum estudo avaliou a utilização do swab vaginal. Objetivos:Avaliar o desempenho do kit AMPLICOR CT\NG (Roche) no diagnóstico de C. trachomatis e N. gonorrhoeae empregando urina, swabs endocervical e vaginal e analisar a concordância de resultados entre as diferentes amostras biológicas. Métodos: A população alvo foi constituída de adolescentes e jovens sexualmente ativas, com idade entre 15 e 24 anos, residentes em Inhumas, Goiás. Os dados sócio-demográficos e de comportamento sexual foram obtidos através de entrevista. O diagnóstico foi realizado empregando o kit Amplicor CT/NG (Roche) em amostras de urina, swab endocervical (SE) e swab vaginal (SV). Para avaliação do desempenho foram calculadas a sensibilidade, especificidade, valor preditivo positivo e negativo dos testes. O coeficiente kappa foi calculado para avaliar a concordância entre as amostras. Considerou-se um resultado como verdadeiro-positivo quando pelo menos duas das três amostras biológicas da mesma paciente fossem positivas para clamídia e/ou gonococo. Resultados: Entre as 428 participantes a média de idade foi de 19,4 anos. Os três espécimes biológicos foram coletados de 309 adolescentes (72,2%). Entre estas, as prevalências foram de 8,7% (IC95% 5,8-12,4)para C. trachomatis e de 2,3% (IC95% 0,9-4,6) para N. gonorrhoeae. Para clamídia as sensibilidades observadas com as diferentes amostras foram superiores a 80% e as especificidades superiores a 97%, com valores preditivos positivos (VPPs) entre 78,8% e 84,6% e valores preditivos negativos (VPNs) >98%. Para o gonococo as sensibilidades foram de 42,8% na urina, 71,4% no SE e de 100% no SV com especificidades >96% nas três amostras. Os dois tipos de swab apresentaram baixos VPPs para o gonococo (≈ 40%) e a urina apresentou VPP de 100%. Os VPNs foram >98%. A concordância de resultados da PCR empregando as diferentes amostras foi de cerca de 94% para ambas as infecções. Entretanto, os valores do coeficiente kappa (κ) variaram de 0,68 a 0,73 para clamídia, o que significa concordância substancial entre as amostras. Para a infecção gonocócica, a concordância foi fraca ou razoável com valores de κ variando de 0,13 a 0,33. Conclusões: Os desempenhos das amostras e os valores do κ sugerem que o swab vaginal parece ser equivalente à urina e ao swab endocervical na detecção da clamídia podendo ser recomendado para estudos de triagem. As três amostras diferiram quanto ao desempenho na detecção da infecção gonocócica e não apresentaram boa concordância de resultados sugerindo que não são equivalentes no diagnóstico desta infecção com o kit de PCR utilizado.

Made available in DSpace on 2014-07-29T15:30:37Z (GMT). No. of bitstreams: 1 Dissertacao Jannaina Karlla de Queiroz Duarte.pdf: 1435551 bytes, checksum: 9764e4445592cad24997ac8311dc77d7 (MD5) Previous issue date: 2008-08-29 / BACKGROUND: Adolescents are at great risk for genital Neisseria gonorrhoeae infection. Although most infections are asymptomatic it can cause severe reproductive sequels to women. Few studies exist about prevalence of these infection in Brazil and fewer in adolescent population. OBJECTIVES: To determine the prevalence of Neisseria gonorrhoeae genital infection among female sexually active adolescents, in Distrito Sanitário Noroeste from Goiânia, Brazil and to identify demographic and behavioral profiles associated to gonococcal infection. METHODS: We performed a cross-sectional study among 427 sexually active female adolescents (15-19 years), random selected at Distrito Sanitário Noroeste and served by the Health Family Program. It were excluded pregnant adolescents, pos-partum, using antibiotics or on period. The gonococcal infection was determined by using polimerase chain reaction (PCR) of Amplicor Roche kit for N. gonorrhoeae and C. trachomatis applied to endocervical swab specimens. Sociodemographic and behavioral data were assessed by face-to-face questionnaire. RESULTS: The mean age of 427 random selected adolescents was 17.2 ± 1.3 years, most female were single (67.9%). The mean age of menarch was 12.4 years and at first sexual intercourse was 15 years. 35,1% had the first sexual intercourse before 15 years and 16,2% reported more than 3 partners in life. About 80% of the participants reported inconsistent condom use. Among those with gonococcal infection, 22,2% also had chlamydial infection. The prevalence of gonococcal infection was 2,1% (95% CI 1,0 4,1). CONCLUSIONS: Adolescents studied presented risk sexual behaviors to sexually transmitted diseases. These results demonstrate the need for preventive medicine programs for adolescents females with ongoing STI prevention activities. / INTRODUÇÃO: Os adolescentes constituem o grupo de maior risco para a infecção genital por Neisseria gonorrhoeae. Embora a maioria das infecções sejam assintomáticas podem causar graves seqüelas reprodutivas nas mulheres. Existem poucos estudos sobre a prevalência desta infecção no Brasil e menos ainda neste grupo populacional. OBJETIVOS: Determinar a prevalência da infecção genital por N. gonorrhoeae em adolescentes sexualmente ativas residentes no Distrito Sanitário Noroeste do município de Goiânia e identificar o perfil sócio-demográfico e comportamental das adolescentes estudadas. METODOLOGIA: Foi realizado um estudo de corte transversal em 427 adolescentes do gênero feminino com idade entre 15 e 19 anos, sexualmente ativas, selecionadas aleatoriamente no Distrito Sanitário Noroeste e atendidas pelo Programa de Saúde da Família. Foram excluídas adolescentes grávidas ou no puerpério, em uso de antibióticos ou menstruadas. O diagnóstico da infecção gonocócica foi realizado empregando a Reação em Cadeia da Polimerase (PCR) em amostras endocervicais utilizando o kit Amplicor Roche para N. gonorrhoeae e C. trachomatis. Foram também obtidas informações sóciodemográficas e de comportamento sexual através de questionário na forma de entrevista. RESULTADOS: A média de idade das 427 adolescentes selecionadas foi de 17,2 ± 1,3 anos, sendo a maioria delas solteira (67,9%). A média de idade da primeira relação sexual foi de 15 anos. 35,1% das adolescentes tiveram a primeira relação sexual antes dos 15 anos e 16,2% relataram mais que três parceiros durante a vida. Cerca de 80% relataram uso inconsistente de preservativo. Dentre aquelas com infecção gonocócica, 22,2% tinham também infecção clamidial. A prevalência encontrada para a infecção gonocócica foi de 2,1% (IC 95% 1,0 4,1). CONCLUSÕES: As adolescentes estudadas apresentaram comportamentos sexuais de risco para a aquisição de DST. Estes dados mostram a necessidade de programas de medicina preventiva direcionados à adolescentes, principalmente atividades de prevenção de DST.

Bakterier har alltid haft en stor inverkan på mänskligheten. För att diagnostisera bakteriella sjukdomar och behandla dem krävs identifiering av bakterien eller bakteriens relevanta egenskaper. Transportmedium har utvecklats för att hålla bakterierna vid liv från provtagning till analys. Syftet med studien var att utvärdera bakteriers viabilitet i det vätskebaserade mediet Copan Eswab jämfört med kolmedium (Copan swab). Bakterierna som ingick i studien var Campylobacter jejuni, Streptococcus pneumoniae, Haemophilus influenzae, Niesseria gonorrhoeae och Fusobacterium nucleatum. Förutom jämförande mellan medierna genomfördes en jämförelse mellan Eswab i kyl och i rumstemperatur. Resultaten för H. influenzae (n=9) och N. gonorrhoeae (n=9) visade att Eswab gav lika många eller fler överlevande bakterier. Gällande F. nucleatum (n=9) visade resultaten att fler överlevde i Copan swab (Copanpinnar) de första 28 timmarna, men även att bakterien inte klarar mer än 28 timmar i rumstemperatur. Gällande S. pneumoniae (n=9) och C. jejuni (n=9) gav båda opålitliga svar. Ytterligare mätpunkter och studier krävs för att erhålla mer pålitliga resultat gällande hur länge bakterierna överlever i Eswab. / Bacteria have always had a great influence on mankind. To diagnose any bacterial disease and treat it it’s necessary to identify the bacteria or any relevant attributes. Different types of specimen transport have been developed to keep the bacteria alive from sampling until the analysis is performed. The purpose of the study was to evaluate the viability of bacteria in the fluid-based media Copan EswabTM compared with charcoal medium (Copan swab). Bacteria included in the study were: Campylobacter jejuni, Streptococcus pneumoniae, Haemophilus influenzae, Niesseria gonorrhoeae and Fusobacterium nucleatum. The study also tried to compare how bacteria survived in Eswab which was refrigerated and in Eswab room temperature. Results for H. influenzae (n=9) and N. gonorrhoeae (n=9) showed that an equal amount or more of the bacteria survived in Eswab. More of F. nucleatum (n=9) survived in Copan swab (Copan swab sticks) for the first 28 hours, additionally they showed that the bacteria won’t survive more than 28 hours in room temperature. Regarding S. pneumoniae (n=9) and C. jejuni (n=9) both displayed unreliable results. Overall more measurements and additional studies are needed for more reliable results.