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1.
PLoS One ; 19(5): e0302400, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38787847

RESUMO

BACKGROUND: In 2012, Botswana introduced 13-valent pneumococcal conjugate vaccine (PCV-13) to its childhood immunization program in a 3+0 schedule, achieving coverage rates of above 90% by 2014. In other settings, PCV introduction has been followed by an increase in carriage or disease caused by non-vaccine serotypes, including some serotypes with a high prevalence of antibiotic resistance. METHODS: We characterized the serotype epidemiology and antibiotic resistance of pneumococcal isolates cultured from nasopharyngeal samples collected from infants (≤12 months) in southeastern Botswana between 2016 and 2019. Capsular serotyping was performed using the Quellung reaction. E-tests were used to determine minimum inhibitory concentrations for common antibiotics. RESULTS: We cultured 264 pneumococcal isolates from samples collected from 150 infants. At the time of sample collection, 81% of infants had received at least one dose of PCV-13 and 53% had completed the three-dose series. PCV-13 serotypes accounted for 27% of isolates, with the most prevalent vaccine serotypes being 19F (n = 20, 8%), 19A (n = 16, 6%), and 6A (n = 10, 4%). The most frequently identified non-vaccine serotypes were 23B (n = 29, 11%), 21 (n = 12, 5%), and 16F (n = 11, 4%). Only three (1%) pneumococcal isolates were resistant to amoxicillin; however, we observed an increasing prevalence of penicillin resistance using the meningitis breakpoint (2016: 41%, 2019: 71%; Cochran-Armitage test for trend, p = 0.0003) and non-susceptibility to trimethoprim-sulfamethoxazole (2016: 55%, 2019: 79%; p = 0.04). Three (1%) isolates were multi-drug resistant. CONCLUSIONS: PCV-13 serotypes accounted for a substantial proportion of isolates colonizing infants in Botswana during a four-year period starting four years after vaccine introduction. A low prevalence of amoxicillin resistance supports its continued use as the first-line agent for non-meningeal pneumococcal infections. The observed increase in penicillin resistance at the meningitis breakpoint and the low prevalence of resistance to ceftriaxone supports use of third-generation cephalosporins for empirical treatment of suspected bacterial meningitis.


Assuntos
Antibacterianos , Testes de Sensibilidade Microbiana , Infecções Pneumocócicas , Vacinas Pneumocócicas , Sorogrupo , Streptococcus pneumoniae , Humanos , Streptococcus pneumoniae/efeitos dos fármacos , Streptococcus pneumoniae/isolamento & purificação , Streptococcus pneumoniae/classificação , Botsuana/epidemiologia , Lactente , Infecções Pneumocócicas/microbiologia , Infecções Pneumocócicas/epidemiologia , Infecções Pneumocócicas/prevenção & controle , Infecções Pneumocócicas/tratamento farmacológico , Vacinas Pneumocócicas/imunologia , Feminino , Antibacterianos/farmacologia , Masculino , Farmacorresistência Bacteriana , Sorotipagem , Nasofaringe/microbiologia , Prevalência
2.
Emerg Infect Dis ; 29(7): 1386-1396, 2023 07.
Artigo em Inglês | MEDLINE | ID: mdl-37308158

RESUMO

Isolating and characterizing emerging SARS-CoV-2 variants is key to understanding virus pathogenesis. In this study, we isolated samples of the SARS-CoV-2 R.1 lineage, categorized as a variant under monitoring by the World Health Organization, and evaluated their sensitivity to neutralizing antibodies and type I interferons. We used convalescent serum samples from persons in Canada infected either with ancestral virus (wave 1) or the B.1.1.7 (Alpha) variant of concern (wave 3) for testing neutralization sensitivity. The R.1 isolates were potently neutralized by both the wave 1 and wave 3 convalescent serum samples, unlike the B.1.351 (Beta) variant of concern. Of note, the R.1 variant was significantly more resistant to type I interferons (IFN-α/ß) than was the ancestral isolate. Our study demonstrates that the R.1 variant retained sensitivity to neutralizing antibodies but evolved resistance to type I interferons. This critical driving force will influence the trajectory of the pandemic.


Assuntos
COVID-19 , Interferon Tipo I , Humanos , SARS-CoV-2/genética , Interferon Tipo I/genética , Anticorpos Neutralizantes , Soroterapia para COVID-19 , Canadá/epidemiologia , Anticorpos Antivirais , Glicoproteína da Espícula de Coronavírus
3.
Trials ; 21(1): 941, 2020 Nov 23.
Artigo em Inglês | MEDLINE | ID: mdl-33225995

RESUMO

OBJECTIVES: 1. To compare the effectiveness of four different surveillance strategies in detecting COVID-19 within the homeless shelter population. 2. To assess the participant adherence over time for each surveillance method. TRIAL DESIGN: This is a prospective cluster-randomized study to compare the effectiveness of four different surveillance regimens across eight homeless shelters in the city of Hamilton. PARTICIPANTS: Participants will include both residents of, and the staff working within, the homeless shelters. All participants aged 18 or older who consent to the study and are able to collect a swab sample (where relevant) are eligible for the study. The study will take place across eight homeless shelters (four men-only and four women-only) in the City of Hamilton in Ontario, Canada. INTERVENTION AND COMPARATOR GROUPS: The comparator group will receive active daily surveillance of symptoms and testing will only be completed in symptomatic participants (i.e. those who fail screening or who seek care for potential COVID-19 related symptoms). The three intervention arms will all receive active daily surveillance of symptoms and testing of symptomatic participants (as in the comparator group) in addition to one of the following: 1. Once weekly self-collected oral swabs (OS) regardless of symptoms using written and visual instructions. 2. Once weekly self-collected oral-nares swab (O-NS) regardless of symptoms using written and visual instructions. 3. Once weekly nurse collected nasopharyngeal swab (NPS) regardless of symptoms. Participants will follow verbal and written instructions for the collection of OS and O-NS specimens. For OS collection, participants are instructed to first moisten the swab on their tongue, insert the swab between the cheek and the lower gums and rotate the swab three times. This is repeated on the other side. For O-NS collection, after oral collection, the swab is inserted comfortably (about 2-3 cm) into one nostril, parallel to the floor and turned three times, then repeated in the other nostril. NPS specimens were collected by the nurse following standard of care procedure. All swabs were placed into a viral inactivation medium and transported to the laboratory for COVID-19 testing. Briefly, total nucleic acid was extracted from specimens and then amplified by RT-PCR for the UTR and Envelope genes of SARS-CoV-2 and the human RNase P gene, which is used as a sample adequacy marker. MAIN OUTCOMES: 1. PRIMARY OUTCOME: COVID-19 detection rate, i.e. the number of new positive cases over the study period of 8 weeks in each arm of the study. 2. SECONDARY OUTCOMES: Qualitative assessment of study enrollment over 8 weeks. Percentage of participants who performed 50% or more of the weekly swabs in the intervention arms in the 8 week study period. RANDOMIZATION: We will use a computer-generated random assignment list to randomize the shelters to one of four interventions. Shelters were stratified by gender, and the simple randomization scheme was applied within each stratum. The randomization scheme was created using WinPEPI. BLINDING: This is an open-label study in which neither participants nor assessors are blinded. NUMBERS TO BE RANDOMIZED (SAMPLE SIZE): Since we are including our total sample frame, a sample size estimation at the cluster level is not required. However, if we succeed to enroll 50 participants per shelter from 8 shelters (n=400), and the detection rate is 3 times higher in the intervention groups (0.15) than in the comparator groups (0.05), we will have 90% power to detect a statistically significant and clinically important difference at a type I error rate of alpha=0.05 (one tailed), assuming an intraclass correlation of ~0.008. These computations were done using WinPEPI, and informed by conservative estimates from other studies on respiratory illness in the homeless (see Full protocol). TRIAL STATUS: The protocol version number is 3.0. Recruitment began on April 17, 2020 and is ongoing. Due to low numbers of COVID cases in the community and shelter system during the initial study period, the trial was extended. The estimated date for the end of the extended recruitment period is Feb 1, 2021. TRIAL REGISTRATION: The trial was registered with ClinicalTrials.gov on June 18, 2020 with the identifier NCT04438070 . FULL PROTOCOL: The full protocol is attached as an additional file, accessible from the Trials website (Additional file 1). In the interest in expediting dissemination of this material, the familiar formatting has been eliminated; this Letter serves as a summary of the key elements of the full protocol.


Assuntos
Infecções por Coronavirus/diagnóstico , Infecções por Coronavirus/prevenção & controle , Pessoas Mal Alojadas/estatística & dados numéricos , Programas de Rastreamento/métodos , Pandemias/prevenção & controle , Pneumonia Viral/diagnóstico , Pneumonia Viral/prevenção & controle , Adulto , Betacoronavirus/genética , COVID-19 , Infecções por Coronavirus/epidemiologia , Infecções por Coronavirus/virologia , Diagnóstico Precoce , Feminino , Humanos , Masculino , Ontário/epidemiologia , Cooperação do Paciente , Pneumonia Viral/epidemiologia , Pneumonia Viral/virologia , Avaliação de Programas e Projetos de Saúde , Estudos Prospectivos , SARS-CoV-2 , Manejo de Espécimes/métodos , Fatores de Tempo
4.
Papillomavirus Res ; 3: 155-159, 2017 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-28720450

RESUMO

SurePath specimens from women referred to colposcopy were treated with Aptima Transfer Solution (ATS) before testing in Aptima HPV (AHPV) and Aptima HPV 16, 18/45 (AHPV-GT) assays. Untreated SurePath specimens were tested with the cobas HPV test. PreservCyt specimens were assessed for cytology and tested with AHPV. High-grade cervical intraepithelial neoplasia lesions served as the reference standard. Excellent agreement (95.5%; k=0.91) was observed for ATS-treated SurePath specimens between Tigris and Panther systems and between the PreservCyt and ATS-treated SurePath specimens (91.1%, k=0.81) with the AHPV assay on Tigris. Agreement between the AHPV and cobas assays with SurePath specimens was substantial (89.9%, k=0.80). AHPV sensitivity for CIN2+(n=147) was 91.2% for SurePath and PreservCyt. Cobas HPV sensitivity was 93.9% for SurePath specimens. AHPV testing of SurePath specimens was more specific (59.4%) than cobas (54.7%) (p<0.001). Detection and genotyping showed similar absolute and relative risks. ATS-treated SurePath specimens tested with AHPV and AHPV-GT assays showed similar performance with greater specificity than cobas HPV on SurePath specimens. Similar overall results were seen using a CIN3 disease endpoint.

5.
Sex Transm Dis ; 44(3): 161-165, 2017 03.
Artigo em Inglês | MEDLINE | ID: mdl-28182591

RESUMO

OBJECTIVES: North American and European advisory groups recommend testing for Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (NG) with nucleic acid amplification tests. Testing is often performed on automated instruments. The objectives of this study were to process urines for the diagnosis of CT and NG and to examine workflow procedures and outcomes. METHODS: While processing 1, 24, 48, 96, and 192 urine specimens on 3 batch-mode systems which use 96-well plates: cobas 4800, m2000, and Viper XTR and the random access cartridge testing GeneXpert Infinity 80, we measured assay performance, hands-on time for processing and maintenance, reagents and plastics consumption, time required to obtain results, and testing accuracy. RESULTS: The Infinity 80 required the least hands-on time for single specimens and smaller batches, whereas the Viper XTR and m2000 required the most hands-on time for all batch sizes. Cumulative daily, weekly, and monthly maintenance was highest for the Viper XTR and lowest for Infinity 80. All batch-mode instruments consumed large amounts of disposables. Time to results was shortest for the Infinity 80, and the Viper XTR provided the shortest time for the batch-mode instruments. All systems showed similar diagnostic accuracy. CONCLUSIONS: Because detection performances were similar, issues of hands-on time, maintenance, time to results, and consumables are important operational factors for the diagnosis and treatment of CT/NG infections.


Assuntos
Infecções por Chlamydia/diagnóstico , Gonorreia/diagnóstico , Técnicas de Amplificação de Ácido Nucleico/instrumentação , Avaliação de Processos e Resultados em Cuidados de Saúde , Manejo de Espécimes/instrumentação , Urinálise/instrumentação , Infecções por Chlamydia/microbiologia , Chlamydia trachomatis , Gonorreia/microbiologia , Humanos , Neisseria gonorrhoeae , Técnicas de Amplificação de Ácido Nucleico/métodos , Sensibilidade e Especificidade , Manejo de Espécimes/métodos , Urinálise/métodos
6.
Sex Transm Dis ; 43(6): 377-81, 2016 06.
Artigo em Inglês | MEDLINE | ID: mdl-27196259

RESUMO

BACKGROUND: The 2015 Sexually Transmitted Diseases Treatment Guidelines from the Centers for Disease Control and Prevention recommend testing for Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (NG) using nucleic acid amplification tests, and prompt treatment of infected persons on site under direct observation. Faster time to results may enable treatment and management outcomes. METHODS: Workflow parameters for processing 1, 10, 48, 96, and 192 tests were determined in the GeneXpert Infinity 80 (Cepheid) and Panther (Hologic) instruments. RESULTS: In an Xpert CT/NG cartridge, the time to first results on the Infinity 80 was 1 hour 30 minutes for single or multiple tests and final results for 10, 48, 96, and 192 tests were available at 1 hour 37 minutes, 1 hour 54 minutes, 3 hour 17 minutes, and 5 hour 7 minutes, respectively. With the Aptima CT/GC assay on the Panther, the respective times were 3 hr 45 min for the first test result, and 3 hour 51 minutes, 4 hour 38 minutes, 5 hour 26 minutes, and 7 hour 4 minutes to final results. The Panther required more time for maintenance and consumed a greater variety of plastics and reagents but required less hands-on time when testing larger numbers of specimens. CONCLUSIONS: The Infinity 80 is a versatile instrument for continuous random access testing of small or large numbers of clinical specimens and may provide diagnostic results, in some settings, in time for treatment of CT and NG infections.


Assuntos
Infecções por Chlamydia/diagnóstico , Chlamydia trachomatis/isolamento & purificação , Gonorreia/diagnóstico , Neisseria gonorrhoeae/isolamento & purificação , Técnicas de Amplificação de Ácido Nucleico/instrumentação , Doenças Bacterianas Sexualmente Transmissíveis/diagnóstico , Infecções por Chlamydia/microbiologia , Chlamydia trachomatis/genética , Gonorreia/microbiologia , Humanos , Neisseria gonorrhoeae/genética , Sensibilidade e Especificidade , Doenças Bacterianas Sexualmente Transmissíveis/microbiologia , Fatores de Tempo , Fluxo de Trabalho
7.
Sex Transm Dis ; 43(2): 125-9, 2016 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-26760183

RESUMO

BACKGROUND: Many sexually transmitted diseases are asymptomatic in the lower genital tract and can cause upper tract complications if left untreated. Self-collected vaginal (SCV) swabs enable the accurate detection of many sexually transmitted infections and give women the option of collecting their own samples while providing them with privacy and convenience. METHODS: We compared SCV samples collected and transported dry using the HerSwab device to physician-collected vaginal (PCV) Aptima swabs for the detection of Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (NG), and measured patients' ease and comfort with self-collection. A total of 189 women aged 16 to 41 years were consented into the study and answered a standardized anonymized questionnaire regarding self-collection with the HerSwab device. RESULTS: Women reported self-collection with HerSwab to be easy (97.1%) and comfortable (88.3%). They preferred self-collection over physician collection (80.9%) and would consider using HerSwab for self-collection at home (79.7%). Samples of SCV and PCV showed an overall agreement of 94.7% (κ = 0.64) for CT and of 98.4% (κ = 0.56) for NG, and HerSwab collection detected 7 more positive patients than PCV collection. The overall prevalence of infection was 10.6% for CT and 2.6% for NG. CONCLUSION: HerSwab SCV samples are suitable for the diagnosis of CT and NG.


Assuntos
Infecções por Chlamydia/diagnóstico , Chlamydia trachomatis/isolamento & purificação , Gonorreia/diagnóstico , Neisseria gonorrhoeae/isolamento & purificação , Infecções Sexualmente Transmissíveis/diagnóstico , Manejo de Espécimes/instrumentação , Adolescente , Adulto , Infecções por Chlamydia/epidemiologia , Chlamydia trachomatis/genética , Demografia , Feminino , Gonorreia/epidemiologia , Humanos , Neisseria gonorrhoeae/genética , Prevalência , Infecções Sexualmente Transmissíveis/epidemiologia , Manejo de Espécimes/métodos , Inquéritos e Questionários , Vagina/microbiologia , Adulto Jovem
8.
J Clin Microbiol ; 52(10): 3808-9, 2014 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-25122855

RESUMO

Self-collected vaginal Aptima swabs and flocked swabs in Aptima specimen transport medium and ESwabs in ESwab medium detected all 37 Chlamydia trachomatis-infected patients from 287 women tested by the Aptima Combo assay. Prevalence rates of C. trachomatis, Neisseria gonorrhoeae, and dual infection were 12.8%, 3.1%, and 2.4%, respectively.


Assuntos
Técnicas Bacteriológicas/métodos , Infecções por Chlamydia/diagnóstico , Chlamydia trachomatis/isolamento & purificação , Gonorreia/diagnóstico , Neisseria gonorrhoeae/isolamento & purificação , Manejo de Espécimes/métodos , Feminino , Humanos , Prevalência
9.
Sex Transm Dis ; 41(6): 365-8, 2014 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-24825332

RESUMO

BACKGROUND: An APTIMA specimen collection and transportation (SCT) kit was developed by Hologic/Gen-Probe. OBJECTIVES: To compare cervical SCT samples to PreservCyt and SurePath samples and self-collected vaginal samples to physician-collected vaginal and cervical SCT samples. To determine ease and comfort of self-collection with the kit. STUDY DESIGN: Each woman (n = 580) self-collected a vaginal SCT, then filled out a questionnaire (n = 563) to determine ease and comfort of self-collection. Colposcopy physicians collected a vaginal SCT and cervical PreservCyt, SCT, and SurePath samples. Samples were tested by APTIMA HPV (AHPV) assay. RESULTS: Agreement between testing of cervical SCT and PreservCyt was 91.1% (κ = 0.82), and that of SurePath samples was 86.7% (κ = 0.72). Agreement of self-collected vaginal SCT to physician-collected SCT was 84.7% (κ = 0.68), and that of self-collected vaginal to cervical SCT was 82.0% (κ = 0.63). For 30 patients with CIN2+, AHPV testing of cervical SCT was 100% sensitive and 59.8% specific compared with PreservCyt (96.6% and 66.2%) and SurePath (93.3% and 70.9%). Vaginal SCT sensitivity was 86.7% for self-collection and 80.0% for physician collection. Most patients found that vaginal self-collection was easy, 5.3% reported some difficulty, and 87.6% expressed no discomfort. CONCLUSIONS: Cervical samples collected with the new SCT kit compared well to traditional liquid-based samples tested by AHPV. Although there was good agreement between self-collected and physician-collected samples with the SCT, in a limited number of 30 women, vaginal sampling identified fewer with CIN2+ precancerous cervical lesions than cervical SCT sampling. Comfort, ease of use, and detection of high-risk HPV demonstrated that the kit could be used for cervical and vaginal sampling.


Assuntos
Colo do Útero/virologia , Papillomaviridae/isolamento & purificação , Infecções por Papillomavirus/patologia , RNA Mensageiro/análise , Displasia do Colo do Útero/patologia , Neoplasias do Colo do Útero/patologia , Esfregaço Vaginal/métodos , Adolescente , Adulto , Colposcopia , Feminino , Humanos , Pessoa de Meia-Idade , Papillomaviridae/genética , Infecções por Papillomavirus/genética , RNA Viral/análise , Kit de Reagentes para Diagnóstico , Reprodutibilidade dos Testes , Autocuidado , Sensibilidade e Especificidade , Manejo de Espécimes/métodos , Neoplasias do Colo do Útero/genética , Displasia do Colo do Útero/genética
10.
J Clin Microbiol ; 52(7): 2299-304, 2014 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-24740081

RESUMO

The choice of a suitable automated system for a diagnostic laboratory depends on various factors. Comparative workflow studies provide quantifiable and objective metrics to determine hands-on time during specimen handling and processing, reagent preparation, return visits and maintenance, and test turnaround time and throughput. Using objective time study techniques, workflow characteristics for processing 96 and 192 tests were determined on m2000 RealTime (Abbott Molecular), Viper XTR (Becton Dickinson), cobas 4800 (Roche Molecular Diagnostics), Tigris (Hologic Gen-Probe), and Panther (Hologic Gen-Probe) platforms using second-generation assays for Chlamydia trachomatis and Neisseria gonorrhoeae. A combination of operational and maintenance steps requiring manual labor showed that Panther had the shortest overall hands-on times and Viper XTR the longest. Both Panther and Tigris showed greater efficiency whether 96 or 192 tests were processed. Viper XTR and Panther had the shortest times to results and m2000 RealTime the longest. Sample preparation and loading time was the shortest for Panther and longest for cobas 4800. Mandatory return visits were required only for m2000 RealTime and cobas 4800 when 96 tests were processed, and both required substantially more hands-on time than the other systems due to increased numbers of return visits when 192 tests were processed. These results show that there are substantial differences in the amount of labor required to operate each system. Assay performance, instrumentation, testing capacity, workflow, maintenance, and reagent costs should be considered in choosing a system.


Assuntos
Automação Laboratorial/métodos , Infecções por Chlamydia/diagnóstico , Testes Diagnósticos de Rotina/métodos , Gonorreia/diagnóstico , Manutenção/métodos , Fluxo de Trabalho , Chlamydia trachomatis/isolamento & purificação , Humanos , Neisseria gonorrhoeae/isolamento & purificação , Fatores de Tempo
11.
J Clin Microbiol ; 52(7): 2305-10, 2014 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-24696024

RESUMO

In a comparison of 4 second-generation nucleic acid amplification tests performed with self-collected vaginal swab (SCVS) and first-void urine (FVU) specimens from 575 women, SCVS specimens indicated more infections than did FVU specimens in all assays. The prevalence rates were 9% (53/575 patients) for Chlamydia trachomatis and 2% (11/575 patients) for Neisseria gonorrhoeae. The clinical sensitivities for testing SCVS specimens for C. trachomatis were 98.1% on a Tigris system and 96.2% on a Panther system for the Aptima Combo 2 assay (Hologic Gen-Probe), 98.0% for the RealTime CT/NG assay on an m2000 instrument (Abbott), 90.6% for the ProbeTec CT/GC Q(x) assay on the Viper system (Becton Dickinson), and 84.6% for the cobas CT/NG assay on the cobas 4800 platform (Roche). Clinical sensitivities for C. trachomatis in FVU specimens were 88.7% (Tigris) and 88.0% (Panther) for the Aptima Combo 2 assay, 76.9% for the RealTime CT/NG assay, 75.5% for the ProbeTec CT/GC Q(x) assay, and 81.1% for the cobas CT/NG assay. Clinical sensitivities of the assays for N. gonorrhoeae, with limited positive results, ranged from 63.6% to 100%. Specificities for both infections ranged from 98.4 to 100%. Differences in analytical sensitivities and levels of molecular targets in clinical samples but not inhibitors of amplification may explain the differences in clinical sensitivities.


Assuntos
Infecções por Chlamydia/diagnóstico , Gonorreia/diagnóstico , Técnicas de Diagnóstico Molecular/métodos , Técnicas de Amplificação de Ácido Nucleico/métodos , Manejo de Espécimes/métodos , Urina/microbiologia , Vagina/microbiologia , Chlamydia trachomatis/isolamento & purificação , Feminino , Cabeça , Humanos , Neisseria gonorrhoeae/isolamento & purificação , Autoadministração , Sensibilidade e Especificidade
12.
J Clin Microbiol ; 52(2): 668-70, 2014 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-24478509

RESUMO

Use of a new collection kit for vaginal and cervical sampling was reported as easy by the majority of 692 women and not uncomfortable (by 87.4% of those ≥ 25 years old and 78.8% of those <25 years old). By Aptima testing, patient- and physician-collected samples agreed strongly for Chlamydia trachomatis (99.6% to 99.3%; κ = 0.93 to 0.89) and T. vaginalis (99.6% to 98.9%; κ = 0.97 to 0.78).


Assuntos
Infecções por Chlamydia/diagnóstico , Chlamydia trachomatis/isolamento & purificação , Manejo de Espécimes/métodos , Meios de Transporte/métodos , Tricomoníase/diagnóstico , Trichomonas vaginalis/isolamento & purificação , Vagina/microbiologia , Vagina/parasitologia , Adolescente , Adulto , Idoso , Infecções por Chlamydia/microbiologia , Feminino , Humanos , Pessoa de Meia-Idade , Tricomoníase/parasitologia , Adulto Jovem
13.
Sex Transm Infect ; 89(2): 102-4, 2013 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-23024224

RESUMO

OBJECTIVES: To compare first catch urine (FCU) and self-collected urinary meatal swabs for the detection of Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (NG) using the APTIMA Combo 2 assay. METHODS: A total of 511 young men from a high risk street youth clinic were studied. Group A (n=293) collected a FCU and a meatal APTIMA swab followed by Group B (n=218) who collected a FCU and two meatal samples using an APTIMA swab and a flocked swab. Order of sample collection was alternated. Individuals in Group B rated collection as easy, difficult or neither, then expressed a preference for sampling and swab type. All subjects performed meatal self-collection in the presence of a study monitor. RESULTS: The combined CT prevalence was 7.8% and 2.7% for NG where 80% of the men were without symptoms. Meatal swabbing identified 35 cases of CT and 14 cases of NG compared to 33 and 11 for FCU. Flocked and APTIMA swabs were equally effective in detecting more cases. The majority of men found self-collection of meatal swabs and urine to be easy. Although 63% preferred urine sampling, 60% of those who preferred swabbing selected the flocked swab. CONCLUSIONS: Collection of meatal swabs could serve as an alternative to urethral swabbing and FCU for the detection of CT and NG.


Assuntos
Infecções por Chlamydia/diagnóstico , Chlamydia trachomatis/isolamento & purificação , Gonorreia/diagnóstico , Neisseria gonorrhoeae/isolamento & purificação , Autoadministração/métodos , Manejo de Espécimes/métodos , Urina/microbiologia , Adolescente , Humanos , Masculino , Adulto Jovem
14.
Sex Transm Infect ; 88(3): 160-2, 2012 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-22166474

RESUMO

OBJECTIVES: To compare self-collected vaginal swab (SCVS) types and first-catch urine (FCU) to diagnose Trichomonas vaginalis using analyte-specific reagents designed to be used in a transcription-mediated amplification assay. METHODS: A total of 241 women (group A) collected a FCU and a SCVS using a dacron swab (APTIMA collection kit). A second group of 289 women (group B) collected two SCVS using one dacron swab and one nylon-flocked swab. RESULTS: Of 75 young women (street youth) determined to be infected with T vaginalis only seven reported symptoms of vaginal discharge or irritation. Using a cutoff of 50,000 relative light units, the sensitivity and specificity was 97.2% and 97.6%, respectively for dacron SCVS compared with 41.7% and 100% for FCU in group A; 92.3% and 98.8% for dacron SCVS and 92.3% and 99.2% for flocked-nylon SCVS in group B. The assay tested 96 samples in 6 h. CONCLUSIONS: Dacron and nylon-flocked SCVS performed equally well and significantly better than FCU using analyte-specific reagents in the APTIMA transcription-mediated amplification assay. Either swab type could be used for self-collection.


Assuntos
DNA de Protozoário/genética , Autoexame/métodos , Manejo de Espécimes/métodos , Vaginite por Trichomonas/diagnóstico , Trichomonas vaginalis/isolamento & purificação , Urina/parasitologia , Vagina/parasitologia , Feminino , Humanos , Técnicas de Diagnóstico Molecular/métodos , Técnicas de Amplificação de Ácido Nucleico/métodos , Nylons , Parasitologia/métodos , Polietilenotereftalatos , Sensibilidade e Especificidade , Vaginite por Trichomonas/parasitologia , Trichomonas vaginalis/genética , Adulto Jovem
15.
J Clin Microbiol ; 49(5): 1970-1, 2011 May.
Artigo em Inglês | MEDLINE | ID: mdl-21411585

RESUMO

Aptima Combo 2 (AC2) Neisseria gonorrhoeae testing of 81,405 patients who were tested by culture and 14,666 who were AC2 tested for Chlamydia trachomatis detected 142 extra infections and confirmed 106 culture-positive samples (the positivity rate increased from 0.13 in testing by culture to 0.26 in testing by AC2). Retrievable AC2 positive samples were confirmed (98.5%) by an alternate AGC test.


Assuntos
Gonorreia/diagnóstico , Técnicas de Diagnóstico Molecular/métodos , Neisseria gonorrhoeae/isolamento & purificação , Infecções por Chlamydia/diagnóstico , Chlamydia trachomatis/genética , Chlamydia trachomatis/isolamento & purificação , Feminino , Humanos , Masculino , Neisseria gonorrhoeae/genética , Técnicas de Amplificação de Ácido Nucleico/métodos , Sensibilidade e Especificidade
16.
BMC Microbiol ; 10: 18, 2010 Jan 22.
Artigo em Inglês | MEDLINE | ID: mdl-20096108

RESUMO

BACKGROUND: Flagellar secretion systems are utilized by a wide variety of bacteria to construct the flagellum, a conserved apparatus that allows for migration towards non-hostile, nutrient rich environments. Chlamydia pneumoniae is an obligate, intracellular pathogen whose genome contains at least three orthologs of flagellar proteins, namely FliI, FlhA and FliF, but the role of these proteins remains unknown. RESULTS: Full length FliI, and fragments of FlhA, FliF, and FliI, were cloned and expressed as either GST or His tagged proteins in E. coli. The GST-tagged full length FliI protein was shown to possess ATPase activity, hydrolyzing ATP at a rate of 0.15 +/- .02 micromol min-1 mg-1 in a time- and dose-dependant manner. Using bacterial-2-hybrid and GST pull-down assays, the N-terminal domain of FliI was shown to interact with the cytoplasmic domain of FlhA, but not with FliF, and the cytoplasmic domain of FlhA was shown to interact with the C-terminus of FliF. The absence of other flagellar orthologs led us to explore cross-reaction of flagellar proteins with type III secretion proteins, and we found that FliI interacted with CdsL and CopN, while FlhA interacted with CdsL and Cpn0322 (YscU ortholog CdsU). CONCLUSIONS: The specific interaction of the four orthologous flagellar proteins in C. pneumoniae suggests that they interact in vivo and, taken together with their conservation across members of the chlamydiae sps., and their interaction with T3S components, suggests a role in bacterial replication and/or intracellular survival.


Assuntos
Proteínas de Bactérias/metabolismo , Chlamydophila pneumoniae/enzimologia , Proteínas de Membrana/metabolismo , Adenosina Trifosfatases/genética , Adenosina Trifosfatases/metabolismo , Sequência de Aminoácidos , Proteínas de Bactérias/genética , Chlamydophila pneumoniae/genética , Glutationa , Proteínas de Membrana/genética , Dados de Sequência Molecular , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismo , Alinhamento de Sequência , Análise de Sequência de DNA , Técnicas do Sistema de Duplo-Híbrido
17.
J Bacteriol ; 190(20): 6580-8, 2008 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-18708502

RESUMO

Type III secretion (T3S) is utilized by a wide range of gram-negative bacterial pathogens to allow the efficient delivery of effector proteins into the host cell cytoplasm through the use of a syringe-like injectisome. Chlamydophila pneumoniae is a gram-negative, obligate intracellular pathogen that has the structural genes coding for a T3S system, but the functionality of the system has not yet been demonstrated. T3S is dependent on ATPase activity, which catalyzes the unfolding of proteins and the secretion of effector proteins through the injectisome. CdsN (Cpn0707) is predicted to be the T3S ATPase of C. pneumoniae based on sequence similarity to other T3S ATPases. Full-length CdsN and a C-terminal truncation of CdsN were cloned as glutathione S-transferase (GST)-tagged constructs and expressed in Escherichia coli. The GST-tagged C-terminal truncation of CdsN possessed ATPase activity, catalyzing the release of ADP and P(i) from ATP at a rate of 0.55 +/- 0.07 micromol min(-1) mg(-1) in a time- and dose-dependent manner. CdsN formed oligomers and high-molecular-weight multimers, as assessed by formaldehyde fixation and nondenaturing polyacrylamide gel electrophoresis. Using bacterial two-hybrid and GST pull-down assays, CdsN was shown to interact with CdsD, CdsL, CdsQ, and CopN, four putative structural components of the C. pneumoniae T3S system. CdsN also interacted with an unannotated protein, Cpn0706, a putative CdsN chaperone. Interactions between CdsN, CdsD, and CopN represent novel interactions not previously reported for other bacterial T3S systems and may be important in the localization and/or function of the ATPase at the inner membrane of C. pneumoniae.


Assuntos
Adenosina Trifosfatases/genética , Adenosina Trifosfatases/metabolismo , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Chlamydophila pneumoniae/enzimologia , Chlamydophila pneumoniae/genética , Proteínas de Membrana Transportadoras/genética , Proteínas de Membrana Transportadoras/metabolismo , Difosfato de Adenosina/metabolismo , Adenosina Trifosfatases/química , Adenosina Trifosfatases/isolamento & purificação , Trifosfato de Adenosina/metabolismo , Sequência de Aminoácidos , Proteínas de Bactérias/química , Proteínas de Bactérias/isolamento & purificação , Clonagem Molecular , Eletroforese em Gel de Poliacrilamida , Escherichia coli/genética , Expressão Gênica , Cinética , Proteínas de Membrana Transportadoras/química , Proteínas de Membrana Transportadoras/isolamento & purificação , Dados de Sequência Molecular , Peso Molecular , Fósforo/metabolismo , Ligação Proteica , Mapeamento de Interação de Proteínas , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/isolamento & purificação , Proteínas Recombinantes/metabolismo , Alinhamento de Sequência , Deleção de Sequência
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