RESUMO
Molecular identification of the invasion plasmid antigen-H (ipaH) gene has been established as a useful detection mechanism for Shigella spp. The Global Enteric Multicenter Study (GEMS) identified the etiology and burden of moderate-to-severe diarrhea (MSD) in sub-Saharan Africa and south Asia using a case-control study and traditional culture techniques. Here, we used quantitative polymerase chain reaction (qPCR) to identify Shigella spp. in 2,611 stool specimens from GEMS and compared these results to those using culture. Demographic and nutritional characteristics were assessed as possible risk factors. The qPCR identified more cases of shigellosis than culture; however, the distribution of demographic characteristics was similar by both methods. In regression models adjusting for Shigella quantity, age, and site, children who were exclusively breast-fed had significantly lower odds of MSD compared with children who were not breast-fed (odds ratio [OR] = 0.47, 95% confidence interval (CI) = 0.28-0.81). The association between Shigella quantity and MSD increased with age, with a peak in children of 24-35 months of age (OR = 8.2, 95% CI = 4.3-15.7) and the relationship between Shigella quantity and disease was greatest in Bangladesh (OR = 13.2, 95% CI = 7.3-23.8). This study found that qPCR identified more cases of Shigella and age, site, and breast-feeding status were significant risk factors for MSD.
Assuntos
Diarreia/epidemiologia , Diarreia/microbiologia , Disenteria Bacilar/complicações , Disenteria Bacilar/epidemiologia , Shigella/isolamento & purificação , África Subsaariana/epidemiologia , Bangladesh/epidemiologia , Pré-Escolar , Fezes/microbiologia , Feminino , Humanos , Lactente , Recém-Nascido , MasculinoRESUMO
Pathogens in the gastrointestinal tract exist within a vast population of microbes. We examined associations between pathogens and composition of gut microbiota as they relate to Shigella spp./enteroinvasive Escherichia coli infection. We analyzed 3,035 stool specimens (1,735 nondiarrheal and 1,300 moderate-to-severe diarrheal) from the Global Enteric Multicenter Study for 9 enteropathogens. Diarrheal specimens had a higher number of enteropathogens (diarrheal mean 1.4, nondiarrheal mean 0.95; p<0.0001). Rotavirus showed a negative association with Shigella spp. in cases of diarrhea (odds ratio 0.31, 95% CI 0.17-0.55) and had a large combined effect on moderate-to-severe diarrhea (odds ratio 29, 95% CI 3.8-220). In 4 Lactobacillus taxa identified by 16S rRNA gene sequencing, the association between pathogen and disease was decreased, which is consistent with the possibility that Lactobacillus spp. are protective against Shigella spp.-induced diarrhea. Bacterial diversity of gut microbiota was associated with diarrhea status, not high levels of the Shigella spp. ipaH gene.
Assuntos
Disenteria Bacilar/epidemiologia , Disenteria Bacilar/microbiologia , Microbiota , Shigella/genética , Fatores Etários , Biodiversidade , Estudos de Casos e Controles , Pré-Escolar , Países em Desenvolvimento , Diarreia/diagnóstico , Diarreia/epidemiologia , Diarreia/microbiologia , Disenteria Bacilar/diagnóstico , Fezes/microbiologia , Fezes/virologia , Trato Gastrointestinal/microbiologia , Trato Gastrointestinal/virologia , Genes Bacterianos , Humanos , Lactente , Recém-Nascido , Metagenoma , Razão de Chances , RNA Ribossômico 16S/genética , Risco , Índice de Gravidade de Doença , Shigella/classificaçãoRESUMO
BACKGROUND: Diarrheal diseases continue to contribute significantly to morbidity and mortality in infants and young children in developing countries. There is an urgent need to better understand the contributions of novel, potentially uncultured, diarrheal pathogens to severe diarrheal disease, as well as distortions in normal gut microbiota composition that might facilitate severe disease. RESULTS: We use high throughput 16S rRNA gene sequencing to compare fecal microbiota composition in children under five years of age who have been diagnosed with moderate to severe diarrhea (MSD) with the microbiota from diarrhea-free controls. Our study includes 992 children from four low-income countries in West and East Africa, and Southeast Asia. Known pathogens, as well as bacteria currently not considered as important diarrhea-causing pathogens, are positively associated with MSD, and these include Escherichia/Shigella, and Granulicatella species, and Streptococcus mitis/pneumoniae groups. In both cases and controls, there tend to be distinct negative correlations between facultative anaerobic lineages and obligate anaerobic lineages. Overall genus-level microbiota composition exhibit a shift in controls from low to high levels of Prevotella and in MSD cases from high to low levels of Escherichia/Shigella in younger versus older children; however, there was significant variation among many genera by both site and age. CONCLUSIONS: Our findings expand the current understanding of microbiota-associated diarrhea pathogenicity in young children from developing countries. Our findings are necessarily based on correlative analyses and must be further validated through epidemiological and molecular techniques.
Assuntos
Diarreia Infantil/microbiologia , Disenteria/microbiologia , Intestinos/microbiologia , Microbiota/genética , Bangladesh , Sequência de Bases , Estudos de Casos e Controles , Pré-Escolar , Fezes/microbiologia , Feminino , Gâmbia , Humanos , Lactente , Recém-Nascido , Quênia , Masculino , Mali , Tipagem Molecular , Pobreza , RNA Bacteriano/genética , RNA Ribossômico 16S/genéticaRESUMO
Cultivation-based assays combined with PCR or enzyme-linked immunosorbent assay (ELISA)-based methods for finding virulence factors are standard methods for detecting bacterial pathogens in stools; however, with emerging molecular technologies, new methods have become available. The aim of this study was to compare four distinct detection technologies for the identification of pathogens in stools from children under 5 years of age in The Gambia, Mali, Kenya, and Bangladesh. The children were identified, using currently accepted clinical protocols, as either controls or cases with moderate to severe diarrhea. A total of 3,610 stool samples were tested by established clinical culture techniques: 3,179 DNA samples by the Universal Biosensor assay (Ibis Biosciences, Inc.), 1,466 DNA samples by the GoldenGate assay (Illumina), and 1,006 DNA samples by sequencing of 16S rRNA genes. Each method detected different proportions of samples testing positive for each of seven enteric pathogens, enteroaggregative Escherichia coli (EAEC), enterotoxigenic E. coli (ETEC), enteropathogenic E. coli (EPEC), Shigella spp., Campylobacter jejuni, Salmonella enterica, and Aeromonas spp. The comparisons among detection methods included the frequency of positive stool samples and kappa values for making pairwise comparisons. Overall, the standard culture methods detected Shigella spp., EPEC, ETEC, and EAEC in smaller proportions of the samples than either of the methods based on detection of the virulence genes from DNA in whole stools. The GoldenGate method revealed the greatest agreement with the other methods. The agreement among methods was higher in cases than in controls. The new molecular technologies have a high potential for highly sensitive identification of bacterial diarrheal pathogens.
Assuntos
Bactérias/isolamento & purificação , Infecções Bacterianas/diagnóstico , Técnicas Bacteriológicas/métodos , Técnicas Biossensoriais/métodos , Diarreia/microbiologia , Fezes/microbiologia , Técnicas de Diagnóstico Molecular/métodos , Adolescente , Adulto , África , Bactérias/classificação , Infecções Bacterianas/microbiologia , Bangladesh , Criança , Pré-Escolar , Feminino , Humanos , Lactente , Recém-Nascido , Masculino , Pessoa de Meia-Idade , Adulto JovemAssuntos
Salmonella typhi/enzimologia , Salmonella typhi/isolamento & purificação , Febre Tifoide/microbiologia , beta-Lactamases/metabolismo , Antibacterianos/farmacologia , Bangladesh , Pré-Escolar , Humanos , Masculino , Testes de Sensibilidade Microbiana , Salmonella typhi/efeitos dos fármacos , beta-Lactamas/farmacologiaRESUMO
ASSURE® Dengue IgA Rapid Test, an immunochromatographic anti-Dengue IgA Rapid Test based on reverse flow technology, was evaluated using archived sera. The sera were obtained during hospital admission and discharge of 204 patients during 2000 to 2001 dengue outbreak in Bangladesh and 220 negative sera collected in 2009. Based on characterization by reference ELISA (nonstructural protein 1 [NS1] Ag ELISA, immunoglobulin M [IgM]-Cap ELISA, and immunoglobulin G [IgG]-Cap ELISA), 179 (87.7%) patients were positive for dengue infection, and the remaining 245 patients had nondengue febrile illness. The performance of Dengue IgA Rapid Test was compared to reference ELISA. Of 179 dengue-positive sera, 79 (44.1%) were positive by NS1 Ag ELISA, 174 (97.2%) were positive by IgM-Cap ELISA, and 142 (79.3%) were positive by IgG-Cap ELISA. Among 142 IgG-positive cases, 121 (85.2%) patients had shown high level of IgG (PanBio units ≥ 22, equivalent to hemagglutination inhibition (HI) titer ≥ 2560) during hospital admission, indicating secondary infections. Dengue IgA Rapid Test demonstrated 99.4% (178 of 179) sensitivity in diagnosing dengue infection with the ability to detect 100% primary (58 of 58) and 99.2% (120 of 121) secondary infections, and the specificity was found 99.2% (2 of 245). The capability of Dengue IgA Rapid Test in detecting dengue infection in terms of day of illness was comparable to reverse transcriptase polymerase chain reaction and was found better than in-house IgM ELISA. Compared with in-house IgM ELISA, Dengue IgA Rapid Test also detected similar number of dengue virus (DENV) 1, DENV 2, and more DENV 3 and DENV 4 cases. The overall performance thus suggested its usefulness as one of the dengue early diagnostic tools where diagnostic facility is limited.
Assuntos
Anticorpos Antivirais/sangue , Vírus da Dengue/imunologia , Dengue/diagnóstico , Imunoglobulina A/sangue , Kit de Reagentes para Diagnóstico , Antígenos Virais/imunologia , Bangladesh , Cromatografia/métodos , Dengue/imunologia , Dengue/virologia , Ensaio de Imunoadsorção Enzimática , Humanos , Sensibilidade e Especificidade , Fatores de Tempo , Proteínas não Estruturais Virais/imunologiaRESUMO
BACKGROUND: We conducted blood culture surveillance to estimate the incidence of typhoid and paratyphoid fever among urban slum residents in Dhaka, Bangladesh. METHODS: Between January 7, 2003 and January 6, 2004, participants were visited weekly to detect febrile illnesses. Blood cultures were obtained at the clinic from patients with fever (≥38°C). Salmonella isolates were assayed for antimicrobial susceptibility. RESULTS: Forty Salmonella Typhi and eight Salmonella Paratyphi A were isolated from 961 blood cultures. The incidence of typhoid fever was 2.0 episodes/1000 person-years, with a higher incidence in children aged<5 years (10.5/1000 person-years) than in older persons (0.9/1000 person-years) (relative risk=12, 95% confidence interval (CI) 6.3-22.6). The incidence of paratyphoid fever was 0.4/1000 person-years without variation by age group. Sixteen S. Typhi isolates were multidrug-resistant (MDR). All S. Paratyphi isolates were pan-susceptible. The duration of fever among patients with an MDR S. Typhi infection was longer than among patients with non-MDR S. Typhi (16±8 vs. 11±4 days, p=0.02) and S. Paratyphi (10±2 days, p=0.04) infections. CONCLUSIONS: Typhoid fever is more common than paratyphoid fever in the urban Bangladeshi slum; children<5 years old have the highest incidence. Multidrug resistance is common in S. Typhi isolates and is associated with prolonged illness. Strategies for typhoid fever prevention in children aged<5 years in Bangladesh, including immunization, are needed.
Assuntos
Febre Paratifoide/epidemiologia , Febre Tifoide/epidemiologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Bangladesh/epidemiologia , Criança , Pré-Escolar , Farmacorresistência Bacteriana Múltipla , Feminino , Humanos , Incidência , Lactente , Recém-Nascido , Masculino , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Febre Paratifoide/tratamento farmacológico , Febre Paratifoide/microbiologia , Vigilância da População , Áreas de Pobreza , Salmonella paratyphi A/efeitos dos fármacos , Salmonella paratyphi A/isolamento & purificação , Salmonella typhi/efeitos dos fármacos , Salmonella typhi/isolamento & purificação , Febre Tifoide/tratamento farmacológico , Febre Tifoide/microbiologia , População Urbana , Adulto JovemRESUMO
During 1999-2006, 156 isolates of Neisseria meningitidis grew from culture of blood or cerebrospinal fluid at International Centre for Diarrhoeal Disease Research, Bangladesh, in Dhaka, Bangladesh. Serogroup A was the most prevalent strain (97.7%); the rest were serogroup B (2.3%). Most cases of invasive meningococcal disease (88.5%) were identified in 2002-2004 and most (87.5%) occurred in children, teenagers, and young adults, which reflected a community-wide increase in meningococcal disease incidence during this period, which was not recognized previously. All isolates were susceptible to penicillin, ampicillin, chloramphenicol, ciprofloxacin, and ceftriaxone. Cotrimoxazole resistance steadily increased from 50% to 100% during 2002-2006. Resistance to azithromycin emerged in 2002 (5%), increased to 31% in 2004, but isolates in 2005-2006 were susceptible. Information from broader hospital settings and population-based data would precisely assess trends and impact to define strategies for optimal prevention and empiric therapy.
Assuntos
Bacteriemia/microbiologia , Líquido Cefalorraquidiano/microbiologia , Infecções Meningocócicas/epidemiologia , Infecções Meningocócicas/microbiologia , Neisseria meningitidis Sorogrupo A/isolamento & purificação , Adolescente , Adulto , Antibacterianos/farmacologia , Bacteriemia/epidemiologia , Bangladesh/epidemiologia , Criança , Pré-Escolar , Farmacorresistência Bacteriana , Feminino , Humanos , Lactente , Recém-Nascido , Masculino , Pessoa de Meia-Idade , Neisseria meningitidis Sorogrupo A/classificação , Neisseria meningitidis Sorogrupo A/efeitos dos fármacos , Projetos Piloto , Estudos Retrospectivos , Sorotipagem , Adulto JovemAssuntos
Antibacterianos/farmacologia , Ciprofloxacina/farmacologia , Farmacorresistência Bacteriana Múltipla , Salmonella typhi/efeitos dos fármacos , Febre Tifoide/epidemiologia , Bangladesh/epidemiologia , Sangue/microbiologia , Meios de Cultura , Humanos , Testes de Sensibilidade Microbiana , Salmonella typhi/isolamento & purificação , Febre Tifoide/microbiologiaRESUMO
We collected acute-phase serum samples from febrile patients at 2 major hospitals in Dhaka, Bangladesh, during an outbreak of dengue fever in 2001. A total of 18% of dengue-negative patients tested positive for leptospirosis. The case-fatality rate among leptospirosis patients (5%) was higher than among dengue fever patients (1.2%).
Assuntos
Dengue/epidemiologia , Surtos de Doenças , Leptospirose/epidemiologia , Adolescente , Adulto , Bangladesh , Feminino , Humanos , Masculino , Estudos Retrospectivos , Fatores de TempoRESUMO
Dengue fever emerged in Bangladesh in 2000. We tested 225 serum samples from febrile patients and 184 blood donors in 1996 and 1997 for dengue antibodies; 55 (24.4% ) febrile patients had dengue antibodies ( 65.5% with secondary infection pattern), compared with one (0.54%) donor (p <0.001), suggesting that dengue transmission was ongoing well before 1996.