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2.
Emerg Infect Dis ; 29(8): 1708-1711, 2023 08.
Artigo em Inglês | MEDLINE | ID: mdl-37486233

RESUMO

We report extensively drug-resistant (XDR) Shigella sonnei infection in an immunocompromised patient in Texas, USA. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry failed to identify XDR Shigella, but whole-genome sequencing accurately characterized the strain. First-line antimicrobials are not effective against emerging XDR Shigella. Fosfomycin, carbapenems, and tigecycline are potential alternatives.


Assuntos
Anti-Infecciosos , Disenteria Bacilar , Shigella , Humanos , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Anti-Infecciosos/farmacologia , Disenteria Bacilar/diagnóstico , Disenteria Bacilar/tratamento farmacológico , Disenteria Bacilar/epidemiologia , Testes de Sensibilidade Microbiana , Shigella sonnei/genética , Estados Unidos/epidemiologia
3.
PLoS One ; 18(6): e0275353, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37319254

RESUMO

Shigellaa Gram-negative, non-motile bacillus, is the primary causative agent of the infectious disease shigellosis, which kills 1.1 million people worldwideevery year. The children under the age of five are primarily the victims of this disease. This study has been conducted to assess the prevalence of shigellosis through selective plating, biochemical test and conventional PCR assays, where the samples were collected from suspected diarrheoal patients. Invasive plasmid antigen H (ipaH) and O-antigenic rfc gene were used to identify Shigella spp. and S. flexneri respectively. For validation of these identification, PCR product of ipaH gene of a sample (Shigella flexneri MZS 191) has been sequenced and submitted to NCBI database (GenBank accession no- MW774908.1). Further this strain has been used as positive control. Out of 204, around 14.2% (n = 29)(P> 0.01) pediatric diarrheoal cases were screened as shigellosis. Another interesting finding was that most of shigellosis affected children were 7 months to 1 year (P> 0.01).The significance of this study lies in the analyses of the occurrenceand the molecular identification of Shigellaspp. and S. flexneri that can be utilized in improving the accurate identification and the treatment of the most severe and alarming shigellosis.


Assuntos
Disenteria Bacilar , Shigella , Criança , Humanos , Disenteria Bacilar/diagnóstico , Disenteria Bacilar/epidemiologia , Bangladesh/epidemiologia , Shigella/genética , Diarreia/epidemiologia , Diarreia/complicações , Shigella flexneri/genética
4.
Emerg Infect Dis ; 29(7): 1473-1475, 2023 07.
Artigo em Inglês | MEDLINE | ID: mdl-37204919

RESUMO

In Los Angeles, California, USA, persistent, refractory shigellosis was diagnosed in an immunocompetent man who has sex with men. Whole-genome sequencing augmented phenotypic antimicrobial susceptibility testing to comprehensively profile bacterial drug resistance and appropriately guide therapy and clear the infection.


Assuntos
Disenteria Bacilar , Shigella , Masculino , Humanos , Shigella flexneri/genética , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Disenteria Bacilar/diagnóstico , Disenteria Bacilar/tratamento farmacológico , Disenteria Bacilar/epidemiologia , Farmacorresistência Bacteriana , Los Angeles , Testes de Sensibilidade Microbiana
5.
PLoS Negl Trop Dis ; 17(3): e0010450, 2023 03.
Artigo em Inglês | MEDLINE | ID: mdl-36857390

RESUMO

Shigellosis is an increasing cause of gastroenteritis in Australia, with prolonged outbreaks reported in remote Aboriginal and Torres Strait Islander (hereafter "First Nations") communities and among men who have sex with men (MSM) in major cities. To determine associations between Shigella species and demographic and geographic factors, we used multivariate negative binomial regression to analyse national case notifications of shigellosis from 2001 to 2019. Between 2001 and 2019, Australian states and territories reported 18,363 shigellosis cases to the National Notifiable Diseases Surveillance System (NNDSS), of which age, sex and organism information were available for >99% (18,327/18,363) of cases. Of the cases included in our analysis, 42% (7,649/18,327) were S. sonnei, 29% (5,267/18,327) were S. flexneri, 1% (214/18,327) were S. boydii, less than 1% (87/18,327) were S. dysenteriae, and species information was unknown for 28% (5,110/18,327) of cases. Males accounted for 54% (9,843/18,327) of cases, and the highest proportion of cases were in children aged 0-4 years (19%; 3,562/18,327). Crude annual notification rates ranged from 2.2 cases per 100,000 in 2003 and 2011 to 12.4 cases per 100,000 in 2019. Nationally, notification rates increased from 2001 to 2019 with yearly notification rate ratios of 1.04 (95% CI 1.02-1.07) for S. boydii and 1.05 (95% CI 1.04-1.06) for S. sonnei. Children aged 0-4 years had the highest burden of infection for S. flexneri, S. sonnei and S. boydii; and males had a higher notification rate for S. sonnei (notification rate ratio 1.24, 95% CI 1.15-1.33). First Nations Australians were disproportionately affected by shigellosis, with the notification rate in this population peaking in 2018 at 92.1 cases per 100,000 population. Over the study period, we also observed a shift in the testing method used to diagnose shigellosis, with culture independent diagnostic testing (CIDT) increasing from 2014; this also coincided with an increase in notifications of untyped Shigella. This change in testing methodology may have contributed to the observed increase in shigellosis notifications since 2014, with CIDT being more sensitive than culture dependent testing methods. The findings of this study provide important insights into the epidemiological characteristics of shigellosis in Australia, including identification of high-risk groups. This can be used to inform public health prevention and control strategies, such as targeted communication programs in First Nations communities and places with high levels of interaction between young children, such as childcare centres. Our study findings also highlight the implications of culture independent testing on shigellosis surveillance, particularly a reduction in the availability of species level information. This emphasises the continued importance of culture dependant testing for national surveillance of shigellosis.


Assuntos
Disenteria Bacilar , Minorias Sexuais e de Gênero , Shigella , Criança , Masculino , Humanos , Pré-Escolar , Disenteria Bacilar/epidemiologia , Disenteria Bacilar/diagnóstico , Homossexualidade Masculina , Austrália/epidemiologia
6.
J Clin Microbiol ; 61(3): e0165222, 2023 03 23.
Artigo em Inglês | MEDLINE | ID: mdl-36853032

RESUMO

Fecal culture for isolation and identification of Shigella may take days. The BioFire FilmArray Gastrointestinal (GI) panel (bioMérieux, France) is a PCR-based assay that detects enteric pathogens including Shigella/enteroinvasive Escherichia coli (EIEC) in about an hour. The aim of this study was to evaluate the impact of GI panel detection of Shigella in a pediatric emergency department (ED) during an outbreak. Stool samples from children with acute gastroenteritis were tested by the GI panel. Test results were either withheld in preintervention (PRE) or reported to clinicians/families in the postintervention (POST) period. The impact of the GI panel testing on patient management and outcomes was measured. Shigella/EIEC was identified by the GI panel in the PRE (n = 30) and POST (n = 21) phase. The GI panel detected more Shigella infections than did culture; six of 31 (19.4%) Shigella GI panel-positive patients who also had stool cultures were missed by culture. Azithromycin therapy was prescribed for 20% of subjects in the PRE phase and 71.4% of subjects in the POST phase (P < 0.001). Time from the clinical encounter until starting azithromycin therapy was shorter in the POST phase (n = 9), 8.25 h (range, 6.37 to 52.37 h), than in the PRE phase (n = 1), 72 h. Six subjects in the PRE phase visited additional providers compared with one in the POST phase. Prompt diagnosis of shigellosis with the GI panel may provide the opportunity for prompt antimicrobial therapy and avoid additional visits to providers due to early definitive diagnosis. Prompt diagnosis of Shigella at an ED visit may optimize patient management and reduce transmission.


Assuntos
Disenteria Bacilar , Shigella , Humanos , Criança , Azitromicina , Fezes , Disenteria Bacilar/diagnóstico , Disenteria Bacilar/tratamento farmacológico , Disenteria Bacilar/epidemiologia , Escherichia coli , Surtos de Doenças
7.
Isr Med Assoc J ; 25(1): 5-7, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-36718728

RESUMO

BACKGROUND: Clinical dysentery causes hundreds of thousands of deaths annually worldwide. However, current recommendations reserve antibiotics for those either clinically sick or with highly suspected cases of shigellosis. This treatment stems from rising antibiotic resistance. Children diagnosed with clinical dysentery in the pediatric emergency department (PED) are regarded more cautiously. OBJECTIVES: To explore the use of antibiotics in children diagnosed with clinical dysentery in the PED. METHODS: A retrospective case study of children with clinical dysentery at a single PED during the years 2015 and 2018. Demographics as well as clinical findings were compared to culture results and antibiotic treatment. RESULTS: The study included 281 children who were diagnosed with clinical dysentery during the study period; 234 (83%) were treated with antibiotics. However, cultures were positive in only 162 cases (58%). Only 32% were Shigella spp. Younger age, fever, and leukocytosis were related to antibiotic treatment. CONCLUSIONS: The diagnosis of clinical dysentery is misgiven commonly in the PED leading to widespread use of antibiotics when not indicated. This treatment may impact antibiotic resistance patterns. Further studies and interventions are necessary to create clear guidelines in the PED setting.


Assuntos
Disenteria Bacilar , Disenteria , Criança , Humanos , Lactente , Antibacterianos/uso terapêutico , Estudos Retrospectivos , Disenteria/tratamento farmacológico , Disenteria Bacilar/diagnóstico , Disenteria Bacilar/tratamento farmacológico , Serviço Hospitalar de Emergência
8.
Zhonghua Yu Fang Yi Xue Za Zhi ; 56(10): 1472-1477, 2022 Oct 06.
Artigo em Chinês | MEDLINE | ID: mdl-36274616

RESUMO

Objective: To understand the epidemiological characteristics of bacillary dysentery with multiple-onset in Henan province from 2005 to 2020. Methods: The reported cases of bacillary dysentery (including confirmed cases and clinically diagnosed cases) in Henan Province from January 2005 to December 2020 were collected through China's National Disease Supervision Information Management System. The main information included gender, age, home address, date of onset and date of diagnosis. The interval between two episodes of the same case was more than 15 days, which was judged as two episodes. The incidence characteristics of bacillary dysentery patients with two or more cases in Henan Province from 2005 to 2020 were analyzed, and the regional distribution map of cases was drawn using ArcGIS software. Results: From 2005 to 2020, a total of 250 430 cases of bacillary dysentery were reported in Henan Province, with a cumulative incidence rate of 228.66/100 000. There were 2 342 cases with two or more attacks. The incidence of recurrent cases of bacillary dysentery increased year by year (χ2trend=2 932.28, P<0.001). There was no significant difference in the incidence of two or more cases of different sexes (χ2=0.39, P=0.540). There was significant difference in the incidence among different age groups (χ2=438.40, P<0.001). The incidence of two or more cases in the 60-69 age group was relatively high (1.70%). The shortest time interval between the onset of the disease was 16 days, and the longest was 5 579 days, with M (Q1, Q3) about 428 (237, 843) days. Compared with healthy people, those with a history of bacterial diseases had a higher risk of developing bacillary dysentery (RR: 4.12, 95%CI: 3.95‒4.29). Conclusion: The proportion of patients with multiple-onset shows an increasing trend, and there is an age difference.


Assuntos
Disenteria Bacilar , Humanos , Disenteria Bacilar/epidemiologia , Disenteria Bacilar/diagnóstico , Incidência
9.
Trop Biomed ; 39(2): 252-256, 2022 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-35838099

RESUMO

Conventional culture method and biochemical tests remain as the 'gold standard' method for the identification of S. sonnei which are time-consuming. We have discovered previously the potential of three OMPs of S. sonnei (33.3 kDa, 43.8 kDa and 100.3 kDa) as biomarkers in the diagnostic test for shigellosis. Here, we evaluated the performance of the outer membrane proteins of S. sonnei for the development of an antibody-based immunoassay for the detection of S. sonnei infections. All threetarget proteins were specifically recognized when probed with S. sonnei sera. In addition, another two potential proteins of molecular weight 29.0 kDa and 88.2 kDa in size were also exclusively recognized by the IgA when probed with S. sonnei sera. The optimized ELISA demonstrated higher sensitivity and specificity which exceeded 86.0%. In conclusion, the identified target proteins showed great potential as diagnostic biomarkers for the detection of S. sonnei infections in patients.


Assuntos
Disenteria Bacilar , Shigella sonnei , Anticorpos Antibacterianos , Disenteria Bacilar/diagnóstico , Ensaio de Imunoadsorção Enzimática , Humanos , Proteínas de Membrana
10.
PLoS Negl Trop Dis ; 16(2): e0010192, 2022 02.
Artigo em Inglês | MEDLINE | ID: mdl-35130310

RESUMO

Understanding the global burden of enterotoxigenic E. coli (ETEC) and Shigella diarrhea as well as estimating the cost effectiveness of vaccines to control these two significant pathogens have been hindered by the lack of a diagnostic test that is rapid, simple, sensitive, and can be applied to the endemic countries. We previously developed a simple and rapid assay, Rapid Loop mediated isothermal amplification based Diagnostic Test (RLDT) for the detection of ETEC and Shigella spp. (Shigella). In this study, the RLDT assay was evaluated in comparison with quantitative PCR (qPCR), culture and conventional PCR for the detection of ETEC and Shigella. This validation was performed using previously collected stool samples from endemic countries, from the travelers to the endemic countries, as well as samples from a controlled human infection model study of ETEC. The performance of RLDT from dried stool spots was also validated. RLDT resulted in excellent sensitivity and specificity compared to qPCR (99% and 99.2% respectively) ranging from 92.3 to 100% for the individual toxin genes of ETEC and 100% for Shigella. Culture was less sensitive compared to RLDT. No significant differences were noted in the performance of RLDT using samples from various sources or stool samples from moderate to severe diarrhea or asymptomatic infections. RLDT performed equally well in detection of ETEC and Shigella from the dried stool samples on filter papers. This study established that RLDT is sufficiently sensitive and specific to be used as a simple and rapid diagnostic assay to detect ETEC and Shigella in endemic countries to determine disease burden of these pathogens in the national and subnational levels. This information will be important to guide public health and policy makers to prioritize resources for accelerating the development and introduction of effective preventative and/or treatment interventions against these enteric infections.


Assuntos
Disenteria Bacilar/diagnóstico , Escherichia coli Enterotoxigênica/isolamento & purificação , Infecções por Escherichia coli/diagnóstico , Técnicas de Diagnóstico Molecular/métodos , Técnicas de Amplificação de Ácido Nucleico/métodos , Shigella/isolamento & purificação , Escherichia coli Enterotoxigênica/genética , Fezes/microbiologia , Humanos , Reação em Cadeia da Polimerase em Tempo Real/métodos , Sensibilidade e Especificidade , Shigella/genética
11.
PLoS Negl Trop Dis ; 16(1): e0010180, 2022 01.
Artigo em Inglês | MEDLINE | ID: mdl-35089927

RESUMO

Enterotoxigenic E. coli (ETEC) and Shigella spp (Shigella) are complex pathogens. The diagnostic assays currently used to detect these pathogens are elaborate or complicated, which make them difficult to apply in resource poor settings where these diseases are endemic. The culture methods used to detect Shigella are not sensitive, and the methods used to detect ETEC are only available in a few research labs. To address this gap, we developed a rapid and simple diagnostic assay-"Rapid LAMP based Diagnostic Test (RLDT)." The six minutes sample preparation method directly from the fecal samples with lyophilized reaction strips and using established Loop-mediated Isothermal Amplification (LAMP) platform, ETEC [heat labile toxin (LT) and heat stable toxins (STh, and STp) genes] and Shigella (ipaH gene) detection was made simple, rapid (<50 minutes), and inexpensive. This assay is cold chain and electricity free. Moreover, RLDT requires minimal equipment. To avoid any end user's bias, a battery-operated, handheld reader was used to read the RLDT results. The results can be read as positive/negative or as real time amplification depending on the end user's need. The performance specifications of the RLDT assay, including analytical sensitivity and specificity, were evaluated using fecal samples spiked with ETEC and Shigella strains. The limit of detection was ~105 CFU/gm of stool for LT, STh, and STp and ~104 CFU/gm of stool for the ipaH gene, which corresponds to about 23 CFU and 1 CFU respectively for ETEC and Shigella per 25uL reaction within 40 minutes. The RLDT assay from stool collection to result is simple, and rapid and at the same time sufficiently sensitive. RLDT has the potential to be applied in resource poor endemic settings for the rapid diagnosis of ETEC and Shigella.


Assuntos
Disenteria Bacilar/diagnóstico , Infecções por Escherichia coli/diagnóstico , Técnicas de Diagnóstico Molecular/métodos , Técnicas de Amplificação de Ácido Nucleico/métodos , DNA Bacteriano/análise , Disenteria Bacilar/microbiologia , Escherichia coli Enterotoxigênica/genética , Escherichia coli Enterotoxigênica/isolamento & purificação , Infecções por Escherichia coli/microbiologia , Fezes/microbiologia , Humanos , Técnicas de Diagnóstico Molecular/instrumentação , Técnicas de Amplificação de Ácido Nucleico/instrumentação , Shigella/genética , Shigella/isolamento & purificação
12.
J Infect Dev Ctries ; 16(12): 1835-1844, 2022 12 31.
Artigo em Inglês | MEDLINE | ID: mdl-36753649

RESUMO

INTRODUCTION: Shigella continues to cause significant morbidity and mortality each year, mostly in under-five children living in developing countries. We investigated the association between Shigella virulence genes and shigellosis. METHODOLOGY: We randomly selected 61 S. flexneri strains isolated from patients in Bangladesh between 2009 and 2013, and evaluated the presence of 140 MDa large-virulence-plasmid (p140), and 22 virulence genes including ipaH, ial, toxin, and T3SS-related genes. RESULTS: We found p140 in 79% (n = 48) and ipaBCD in 90% (n = 55) strains, while seven strains were missing the p140. The prevalence of ial was 89%, ipgC and ipgE was 85%, and the prevalence for the remaining genes was < 85%. During the multivariate analysis, we found that instead of sen, the Shigella enterotoxin gene set along with several other virulence genes such as ipgA, icsB, ipgB1, spa15, and mxiC, were significantly influencing multiple clinical features relevant to shigellosis, including bloody stool, mucoid stool, and rectal straining. CONCLUSIONS: We believe our model will help to determine the actual disease burden by directly looking for the genetic material in clinically suggestive patients, especially when detecting the causative organisms by traditional means is difficult.


Assuntos
Disenteria Bacilar , Shigella , Humanos , Disenteria Bacilar/epidemiologia , Disenteria Bacilar/diagnóstico , Plasmídeos , Shigella/genética , Shigella flexneri/genética , Virulência/genética , Fatores de Virulência/genética
13.
J Clin Microbiol ; 60(1): e0177421, 2022 01 19.
Artigo em Inglês | MEDLINE | ID: mdl-34669456

RESUMO

Molecular diagnostic methods improve the detection of Shigella, yet their ability to detect Shigella drug resistance on direct stool specimens is less clear. We tested 673 stool specimens from a Shigella treatment study in Bangladesh, including 154 culture-positive stool specimens and their paired Shigella isolates. We utilized a TaqMan array card that included quantitative PCR (qPCR) assays for 24 enteropathogens and 36 antimicrobial resistance (AMR) genes. Shigella was detected by culture in 23% of stool specimens (154/673), while qPCR detected Shigella at diarrhea-associated quantities in 49% (329/673; P < 0.05). qPCR for AMR genes on the Shigella isolates yielded >94% sensitivity and specificity compared with the phenotypic susceptibility results for azithromycin and ampicillin. The performance for trimethoprim-sulfamethoxazole susceptibility was less robust, and the assessment of ciprofloxacin was limited because most isolates were resistant. The detection of AMR genes in direct stool specimens generally yielded low specificities for predicting the resistance of the paired isolate, whereas the sensitivity and negative predictive values for predicting susceptibility were often higher. For example, the detection of ermB or mphA in stool yielded a specificity of 56% but a sensitivity of 91% and a negative predictive value of 91% versus the paired isolate's azithromycin resistance result. Patients who received azithromycin prior to presentation were universally culture negative (0/112); however, qPCR still detected Shigella at diarrhea-associated quantities in 34/112 (30%). In sum, molecular diagnostics on direct stool specimens greatly increase the diagnostic yield for Shigella, including in the setting of prior antibiotics. The molecular detection of drug resistance genes in direct stool specimens had low specificity for confirming resistance but could potentially "rule out" macrolide resistance.


Assuntos
Disenteria Bacilar , Shigella , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Farmacorresistência Bacteriana/genética , Disenteria Bacilar/diagnóstico , Disenteria Bacilar/tratamento farmacológico , Fezes , Humanos , Macrolídeos/farmacologia , Testes de Sensibilidade Microbiana , Shigella/genética
14.
Curr Opin Infect Dis ; 34(5): 463-470, 2021 10 01.
Artigo em Inglês | MEDLINE | ID: mdl-34261903

RESUMO

PURPOSE OF REVIEW: To describe the impact of molecular diagnostics on our understanding of the burden and epidemiology of shigellosis in children in low-income and middle-income countries. RECENT FINDINGS: The incorporation of molecular diagnostics has led to a substantial increase in estimates of the burden of shigellosis and have allowed for further resolution of other aspects of Shigella epidemiology, including the clinical characteristics of shigellosis, the association between clinical and subclinical Shigella infection and linear growth shortfalls, protection after natural infection, duration of convalescent shedding, and host determinants of susceptibility. SUMMARY: The increased sensitivity and precision afforded by molecular approaches has represented a major advance in our understanding of the epidemiology and burden of shigellosis in the settings of highest importance.


Assuntos
Disenteria Bacilar , Shigella , Criança , Pré-Escolar , Países em Desenvolvimento , Disenteria Bacilar/diagnóstico , Disenteria Bacilar/epidemiologia , Humanos , Lactente , Patologia Molecular , Shigella/genética
15.
Braz J Microbiol ; 52(4): 1745-1753, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34251610

RESUMO

Historically, clinical microbiological laboratories have often relied on isolation of pure cultures and phenotypic testing to identify microorganisms. These clinical tests are often based on specific biochemical reactions, growth characteristics, colony morphology, and other physiological aspects. The features used for identification in clinical laboratories are highly conserved and specific for a given group of microbes. We speculate that these features might be the result of evolutionary selection and thus may reflect aspects of the life cycle of the organism and pathogenesis. Indeed, several of the metabolic pathways targeted by diagnostic tests in some cases may represent mechanisms for host colonization or pathogenesis. Examples include, but are not restricted to, Staphylococcus aureus, Pseudomonas aeruginosa, Klebsiella pneumoniae, Salmonella enterica, Shigella spp., and enteroinvasive Escherichia coli (EIEC). Here, we provide an overview of how some common tests reflect molecular mechanisms of bacterial pathogenesis.


Assuntos
Infecções Bacterianas , Disenteria Bacilar , Adaptação ao Hospedeiro , Bactérias/imunologia , Infecções Bacterianas/diagnóstico , Infecções Bacterianas/microbiologia , Infecções Bacterianas/patologia , Disenteria Bacilar/diagnóstico , Disenteria Bacilar/microbiologia , Disenteria Bacilar/fisiopatologia , Humanos , Laboratórios Clínicos
16.
Nat Commun ; 12(1): 2684, 2021 05 11.
Artigo em Inglês | MEDLINE | ID: mdl-33976138

RESUMO

Shigella sonnei is the most common agent of shigellosis in high-income countries, and causes a significant disease burden in low- and middle-income countries. Antimicrobial resistance is increasingly common in all settings. Whole genome sequencing (WGS) is increasingly utilised for S. sonnei outbreak investigation and surveillance, but comparison of data between studies and labs is challenging. Here, we present a genomic framework and genotyping scheme for S. sonnei to efficiently identify genotype and resistance determinants from WGS data. The scheme is implemented in the software package Mykrobe and tested on thousands of genomes. Applying this approach to analyse >4,000 S. sonnei isolates sequenced in public health labs in three countries identified several common genotypes associated with increased rates of ciprofloxacin resistance and azithromycin resistance, confirming intercontinental spread of highly-resistant S. sonnei clones and demonstrating the genomic framework can facilitate monitoring the spread of resistant clones, including those that have recently emerged, at local and global scales.


Assuntos
Disenteria Bacilar/diagnóstico , Genoma Bacteriano/genética , Genômica/métodos , Shigella sonnei/genética , Antibacterianos/farmacologia , Austrália , Azitromicina/farmacologia , Ciprofloxacina/farmacologia , Farmacorresistência Bacteriana Múltipla/genética , Disenteria Bacilar/microbiologia , Inglaterra , Genética Populacional , Genótipo , Geografia , Saúde Global , Humanos , Testes de Sensibilidade Microbiana/métodos , Filogenia , Polimorfismo de Nucleotídeo Único , Shigella sonnei/classificação , Shigella sonnei/fisiologia , Estados Unidos , Sequenciamento Completo do Genoma
17.
Clin Lab ; 67(4)2021 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-33865251

RESUMO

BACKGROUND: Shigellosis is a significant public health challenge particularly in developing countries, and the spread of antibiotic resistance genes through integron structures has become an important problem in the treatment of Shigellosis. The aim of this study was to investigate the genetic association of Shigella flexneri antibiotic resistant clones collected from Ahvaz between 2013 and 2015 by pulse field gel electrophoresis (PFGE) method. METHODS: A total of 45 S. flexneri isolates, which were resistant to ampicillin and cotrimoxazole, were obtained from patients with Shigellosis referred to Ahvaz hospitals during 2013 - 2015. PCR was performed to evaluate the frequency of Sul1, int1, blaOXA, and int2 genes. In addition, pulse field gel electrophoresis method was used to investigate the genetic relationship between 40 S. flexneri isolates. RESULTS: PCR results showed that the highest frequency was related to the sul1 gene with 80% (36 isolates) and the lowest frequency was related to class 2 integron with 15.5% (7 isolates); 31.11% (14 isolates) of the isolates were sul1 and int1. Also, 13.33% (6 isolates) had blaOXA and int1 genes, simultaneously. But none of the isolates had class 1 integrons and class 2 integrons at the same time. PFGE results showed 25 different pulsotype patterns, of which 16 isolates had their own unique pattern and were divided into 16 pulsotypes, and 27 isolates were divided into 9 pulsotypes. CONCLUSIONS: int2 and sul1 resistance genes had an upward trend from 2013 to 2015 and the results of PFGE indicated a different origin of S. flexneri clones.


Assuntos
Disenteria Bacilar , Shigella flexneri , Antibacterianos/farmacologia , Disenteria Bacilar/diagnóstico , Disenteria Bacilar/tratamento farmacológico , Humanos , Integrons/genética , Irã (Geográfico) , Testes de Sensibilidade Microbiana , Shigella flexneri/genética
18.
EBioMedicine ; 66: 103310, 2021 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-33862589

RESUMO

BACKGROUND: Shigellosis is a major cause of moderate to severe diarrhoea and dysentery in children under 5 years of age in low and middle-income countries. The Flexyn2a vaccine conjugates the O-polysaccharide of Shigella flexneri 2a to Pseudomonas aeruginosa exotoxin A. We describe a Phase 2b proof-of-concept challenge study that evaluated safety, immunogenicity, and efficacy of the Flexyn2a vaccine to protect against shigellosis. METHODS: In this randomized, double blind, placebo-controlled trial, healthy adults were randomized 1:1 to receive Flexyn2a (10 µg) or placebo intramuscularly, twice, 4 weeks apart, followed by challenge 4 weeks later with 1500 colony forming units (CFUs) of S. flexneri 2a strain 2457T. The primary outcome was vaccine-induced protection. S. flexneri 2a lipopolysaccharide (LPS)-specific immune responses were assessed. FINDINGS: Sixty-seven subjects were enrolled, 34 received vaccine and 33 placebo. The vaccine was well tolerated; the majority of adverse events were mild in nature. Thirty vaccinees and 29 placebo recipients received the S. flexneri 2a challenge. Vaccination resulted in a 30.2% reduction in shigellosis compared with placebo (13/30 vs. 18/29; p = 0.11; 95% CI -15 to 62.6). Vaccine efficacy was more robust against severe disease, reaching 51.7% (p = 0.015, 95% CI 5.3 to 77.9) against moderate/severe diarrhoea or dysentery concurrent with fever or severe enteric symptoms and 72.4% (p = 0.07) against more severe diarrhoea (≥10 lose stools or ≥1000 g loose stools/24 h). Vaccinated subjects were less likely to need early antibiotic intervention following challenge (protective efficacy 51.7%, p = 0.01; 95% CI 9 to 76.8). In those who developed shigellosis, vaccinated subjects had a lower disease severity score (p = 0.002) than placebo-recipients. Additionally, LPS-specific serum IgG responses in Flexyn2a recipients were associated with protection against disease (p = 0.0016) and with a decreased shigellosis disease score (p = 0.002). INTERPRETATION: The Flexyn2a bioconjugate vaccine was immunogenic, well tolerated and protected against severe illness after Shigella challenge and is a promising Shigella vaccine construct. We identified a strong association between anti-S. flexneri 2a serum IgG and a reduction in disease outcomes. (Clinicaltrials.gov, NCT02646371.) FUNDING: Funding for this study was through a grant from the Wellcome Trust.


Assuntos
Disenteria Bacilar/imunologia , Disenteria Bacilar/prevenção & controle , Vacinas contra Shigella/imunologia , Shigella/imunologia , Adulto , Anticorpos Antibacterianos/sangue , Anticorpos Antibacterianos/imunologia , Especificidade de Anticorpos/imunologia , Disenteria Bacilar/diagnóstico , Feminino , Humanos , Imunoglobulina G/sangue , Imunoglobulina G/imunologia , Lipopolissacarídeos/imunologia , Masculino , Pessoa de Meia-Idade , Vacinas contra Shigella/administração & dosagem , Vacinas contra Shigella/efeitos adversos , Shigella flexneri/imunologia , Resultado do Tratamento , Vacinação , Adulto Jovem
19.
Brief Bioinform ; 22(5)2021 09 02.
Artigo em Inglês | MEDLINE | ID: mdl-33758907

RESUMO

MOTIVATION: Volcano plots are used to select the most interesting discoveries when too many discoveries remain after application of Benjamini-Hochberg's procedure (BH). The volcano plot suggests a double filtering procedure that selects features with both small adjusted $P$-value and large estimated effect size. Despite its popularity, this type of selection overlooks the fact that BH does not guarantee error control over filtered subsets of discoveries. Therefore the selected subset of features may include an inflated number of false discoveries. RESULTS: In this paper, we illustrate the substantially inflated type I error rate of volcano plot selection with simulation experiments and RNA-seq data. In particular, we show that the feature with the largest estimated effect is a very likely false positive result. Next, we investigate two alternative approaches for multiple testing with double filtering that do not inflate the false discovery rate. Our procedure is implemented in an interactive web application and is publicly available.


Assuntos
Simulação por Computador , Genômica/métodos , RNA-Seq/métodos , Estudos de Casos e Controles , Criança , Diarreia/sangue , Diarreia/virologia , Disenteria Bacilar/diagnóstico , Disenteria Bacilar/microbiologia , Expressão Gênica , Humanos , Modelos Lineares , Fenótipo , Reprodutibilidade dos Testes , Rotavirus/genética , Infecções por Rotavirus/diagnóstico , Infecções por Rotavirus/virologia , Salmonella/genética , Infecções por Salmonella/diagnóstico , Infecções por Salmonella/microbiologia , Shigella/genética
20.
BMC Microbiol ; 21(1): 6, 2021 01 06.
Artigo em Inglês | MEDLINE | ID: mdl-33407134

RESUMO

BACKGROUND: The widespread distribution of antimicrobial-resistant Shigella has become a recurrent challenge in many parts of the developing world. Previous studies indicate that the host of Shigella has expanded from humans to animals. This study aimed to investigate the prevalence of fluoroquinolone resistance and associated molecular characterization of S. dysenteriae 1 isolated from calves. RESULTS: All 38 unduplicated S. dysenteriae 1 isolates were collected from calves in Gansu Province from October 2014 to December 2016. According to MLST and PFGE analysis, these isolates were separated into 4 and 28 genotypes, respectively. The most common STs identified were ST228 (34.21%, 13/38) and ST229 (39.47%, 15/38), which were first found in the present study. All isolates harbored virulence genes, and the incidence of the seven virulence genes were ipaH (100%), ipaBCD (92.11%), stx (73.68%), ial (57.89%), sen (28.95%), set1A and set1B (0%). According to the results of antimicrobial susceptibilities, 76.32% (29/38) were resistant to fluoroquinolone and showed multidrug resistance. In a study on the polymorphism of quinolone resistance-determining region (QRDR) of gyrA/B and parC/E genes, we identified two mutations in gyrA (Ser83 → Leu and Asp87 → Asn) and parC (Ser80 → Ile and Ser83 → Leu), respectively. Among them, 55.17% (16/29) of resistant strains had the gyrA point mutations (Ser83 → Leu) and parC point mutation (Ser83 → Leu). Moreover, 41.38% (12/29) of isolates had all five point mutations of gyrA and parC. In addition, the prevalence of the plasmid-mediated quinolone resistance (PMQR) determinant genes was also investigated. All 29 fluoroquinolone-resistant isolates were positive for the aac (6')-Ib-cr gene but negative for qepA, except for SD001. In addition, only 6 (20.69%, 6/29) isolates harbored the qnr gene, including two with qnrB (6.90%, 2/29) and four with qnrS (13.79%, 4/29). CONCLUSION: Given the increased common emergence of multidrug resistant isolates, uninterrupted surveillance will be necessary to understand the actual epidemic burden and control this infection.


Assuntos
Doenças dos Bovinos/microbiologia , Farmacorresistência Bacteriana , Disenteria Bacilar/epidemiologia , Disenteria Bacilar/veterinária , Shigella dysenteriae/patogenicidade , Fatores de Virulência/genética , Animais , Proteínas de Bactérias/genética , Bovinos , Doenças dos Bovinos/diagnóstico , Disenteria Bacilar/diagnóstico , Eletroforese em Gel de Campo Pulsado , Fluoroquinolonas/farmacologia , Regulação Bacteriana da Expressão Gênica/efeitos dos fármacos , Genótipo , Tipagem de Sequências Multilocus , Mutação , Plasmídeos/genética , Prevalência , Shigella dysenteriae/classificação , Shigella dysenteriae/efeitos dos fármacos , Shigella dysenteriae/genética
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