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PulseNet International (PNI) is a global network of 88 countries who work together through their regional and national public health laboratories to track foodborne disease around the world. The vision of PNI is to implement globally standardized surveillance using whole genome sequencing (WGS) for real-time identification and subtyping of foodborne pathogens to strengthen preparedness and response and lower the burden of disease. Several countries in North America and Europe have experienced significant benefits in disease mitigation after implementing WGS. To broaden the routine use of WGS around the world, challenges and barriers must be overcome. We conducted this study to determine the challenges and barriers countries are encountering in their attempts to implement WGS and to identify how PNI can provide support to improve and become a better integrated system overall. A survey was designed with a set of qualitative questions to capture the status, challenges, barriers, and successes of countries in the implementation of WGS and was administered to laboratories in Africa, Asia-Pacific, Latin America and the Caribbean, and Middle East. One-third of respondents do not use WGS, and only 8% reported using WGS for routine, real-time surveillance. The main barriers for implementation of WGS were lack of funding, gaps in expertise, and training, especially for data analysis and interpretation. Features of an ideal system to facilitate implementation and global surveillance were identified as an all-in-one software that is free, accessible, standardized and validated. This survey highlights the minimal use of WGS for foodborne disease surveillance outside the United States, Canada, and Europe to date. Although funding remains a major barrier to WGS-based surveillance, critical gaps in expertise and availability of tools must be overcome. Opportunities to seek sustainable funding, provide training, and identify solutions for a globally standardized surveillance platform will accelerate implementation of WGS worldwide.
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Países em Desenvolvimento , Doenças Transmitidas por Alimentos , Surtos de Doenças , Doenças Transmitidas por Alimentos/epidemiologia , Genoma Bacteriano , Humanos , Inquéritos e Questionários , Estados Unidos/epidemiologia , Sequenciamento Completo do GenomaRESUMO
INTRODUCTIONIn November 2015, the Centers for Disease Control and Prevention (CDC) sent a letter to state and territorial epidemiologists, state and territorial public health laboratory directors, and state and territorial health officials. In this letter, culture-independent diagnostic tests (CIDTs) for detection of enteric pathogens were characterized as "a serious and current threat to public health surveillance, particularly for Shiga toxin-producing Escherichia coli (STEC) and Salmonella" The document says CDC and its public health partners are approaching this issue, in part, by "reviewing regulatory authority in public health agencies to require culture isolates or specimen submission if CIDTs are used." Large-scale foodborne outbreaks are a continuing threat to public health, and tracking these outbreaks is an important tool in shortening them and developing strategies to prevent them. It is clear that the use of CIDTs for enteric pathogen detection, including both antigen detection and multiplex nucleic acid amplification techniques, is becoming more widespread. Furthermore, some clinical microbiology laboratories will resist the mandate to require submission of culture isolates, since it will likely not improve patient outcomes but may add significant costs. Specimen submission would be less expensive and time-consuming for clinical laboratories; however, this approach would be burdensome for public health laboratories, since those laboratories would need to perform culture isolation prior to typing. Shari Shea and Kristy Kubota from the Association of Public Health Laboratories, along with state public health laboratory officials from Colorado, Missouri, Tennessee, and Utah, will explain the public health laboratories' perspective on why having access to isolates of enteric pathogens is essential for public health surveillance, detection, and tracking of outbreaks and offer potential workable solutions which will allow them to do this. Marc Couturier of ARUP Laboratories and Melissa Miller of the University of North Carolina will explain the advantages of CIDTs for enteric pathogens and discuss practical solutions for clinical microbiology laboratories to address these public health needs.
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Testes Diagnósticos de Rotina/métodos , Surtos de Doenças , Monitoramento Epidemiológico , Doenças Transmitidas por Alimentos/diagnóstico , Doenças Transmitidas por Alimentos/epidemiologia , Política de Saúde , Centers for Disease Control and Prevention, U.S. , Humanos , Estados UnidosRESUMO
PulseNet International is a global network dedicated to laboratory-based surveillance for food-borne diseases. The network comprises the national and regional laboratory networks of Africa, Asia Pacific, Canada, Europe, Latin America and the Caribbean, the Middle East, and the United States. The PulseNet International vision is the standardised use of whole genome sequencing (WGS) to identify and subtype food-borne bacterial pathogens worldwide, replacing traditional methods to strengthen preparedness and response, reduce global social and economic disease burden, and save lives. To meet the needs of real-time surveillance, the PulseNet International network will standardise subtyping via WGS using whole genome multilocus sequence typing (wgMLST), which delivers sufficiently high resolution and epidemiological concordance, plus unambiguous nomenclature for the purposes of surveillance. Standardised protocols, validation studies, quality control programmes, database and nomenclature development, and training should support the implementation and decentralisation of WGS. Ideally, WGS data collected for surveillance purposes should be publicly available, in real time where possible, respecting data protection policies. WGS data are suitable for surveillance and outbreak purposes and for answering scientific questions pertaining to source attribution, antimicrobial resistance, transmission patterns, and virulence, which will further enable the protection and improvement of public health with respect to food-borne disease.
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Bases de Dados Factuais , Surtos de Doenças , Microbiologia de Alimentos , Doenças Transmitidas por Alimentos/microbiologia , Genoma Bacteriano , Saúde Pública , Sequenciamento Completo do Genoma/normas , Bases de Dados Factuais/normas , Doenças Transmitidas por Alimentos/epidemiologia , Humanos , Laboratórios , Tipagem de Sequências MultilocusRESUMO
Listeria monocytogenes (Lm) causes severe foodborne illness (listeriosis). Previous molecular subtyping methods, such as pulsed-field gel electrophoresis (PFGE), were critical in detecting outbreaks that led to food safety improvements and declining incidence, but PFGE provides limited genetic resolution. A multiagency collaboration began performing real-time, whole-genome sequencing (WGS) on all US Lm isolates from patients, food, and the environment in September 2013, posting sequencing data into a public repository. Compared with the year before the project began, WGS, combined with epidemiologic and product trace-back data, detected more listeriosis clusters and solved more outbreaks (2 outbreaks in pre-WGS year, 5 in WGS year 1, and 9 in year 2). Whole-genome multilocus sequence typing and single nucleotide polymorphism analyses provided equivalent phylogenetic relationships relevant to investigations; results were most useful when interpreted in context of epidemiological data. WGS has transformed listeriosis outbreak surveillance and is being implemented for other foodborne pathogens.
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Surtos de Doenças , Doenças Transmitidas por Alimentos/epidemiologia , Genoma Bacteriano/genética , Listeria monocytogenes/classificação , Listeriose/epidemiologia , Sequenciamento Completo do Genoma/métodos , Inocuidade dos Alimentos , Doenças Transmitidas por Alimentos/microbiologia , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Listeria monocytogenes/genética , Listeria monocytogenes/isolamento & purificação , Listeriose/microbiologia , Tipagem de Sequências Multilocus , Filogenia , Análise de Sequência de DNARESUMO
Over 1,100 foodborne disease outbreaks cause over 23,000 illnesses in the United States annually, but the rates of outbreaks reported and successful investigation vary dramatically among states. We used data from the Centers for Disease Control and Prevention's outbreak reporting database, Association of Public Health Laboratories' PulseNet laboratory subtyping network survey and Salmonella laboratory survey, national public health surveillance data, and national surveys to examine potential causes of this variability. The mean rate of reporting of Salmonella outbreaks was higher in states requiring submission of all isolates to the state public health laboratory, compared to those that do not (5.9 vs. 4.1 per 10 million population, p=0.0062). Rates of overall outbreak reporting or successful identification of an etiology or food vehicle did not correlate at the state level with population, rates of sporadic disease reporting, health department organizational structure, or self-reported laboratory or epidemiologic capacity. Foodborne disease outbreak surveillance systems are complex, and improving them will require a multi-faceted approach to identifying and overcoming barriers.
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Surtos de Doenças/estatística & dados numéricos , Doenças Transmitidas por Alimentos/epidemiologia , Vigilância da População , Infecções por Salmonella/epidemiologia , Coleta de Dados , Microbiologia de Alimentos , Doenças Transmitidas por Alimentos/microbiologia , Humanos , Saúde Pública , Salmonella/fisiologia , Infecções por Salmonella/microbiologia , Estados Unidos/epidemiologiaRESUMO
Background: Pathogen detection has changed with increased use of culture-independent diagnostic tests (CIDTs). CIDTs do not yield isolates, which are necessary to detect outbreaks using whole-genome sequencing. The Foodborne Diseases Active Surveillance Network (FoodNet) monitors clinical laboratory testing practices to improve interpretation of surveillance data and assess availability of isolates. We describe changes in practices over 8 years. Methods: During 2012-2019, 10 FoodNet sites collected standardized data about practices in clinical laboratories (range, 664-723 laboratories) for select enteric pathogens. We assessed changes in practices. Results: During 2012-2019, the percentage of laboratories that used only culture methods decreased, with the largest declines for Vibrio (99%-57%) and Yersinia (99%-60%). During 2019, the percentage of laboratories using only CIDTs was highest for Shiga toxin-producing Escherichia coli (43%), Campylobacter (34%), and Vibrio (34%). From 2015 to 2019, the percentage of laboratories that performed reflex culture after a positive CIDT decreased, with the largest declines for Shigella (75%-42%) and Salmonella (70%-38%). The percentage of laboratories that routinely submitted isolates to a public health laboratory decreased for all bacterial pathogens examined from 2015 to 2019. Conclusions: By increasing use of CIDTs and decreasing reflex culture, clinical laboratories have transferred the burden of isolate recovery to public health laboratories. Until technologies allow for molecular subtyping directly from a patient specimen, state public health laboratories should consider updating enteric disease reporting requirements to include submission of isolates or specimens. Public health laboratories need resources for isolate recovery.
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BACKGROUND: In the United States, tularemia is caused by Francisella tularensis subsps. tularensis (type A) and holarctica (type B). Molecular subtyping has further divided type A into 2 subpopulations, A1 and A2. Significant mortality differences were previously identified between human infections caused by A1 (14%), A2 (0%) and type B (7%). To verify these findings and to further define differences among genotypes, we performed a large-scale molecular epidemiologic analysis of F. tularensis isolates from humans and animals. METHODS: Pulsed-field gel electrophoresis with PmeI was performed on 302 type A and 61 type B isolates. Pulsed-field gel electrophoresis pattern and epidemiologic analyses were performed. Logistic regression was used to assess factors associated with human mortality. RESULTS: Pulsed-field gel electrophoresis typing identified 4 distinct type A genotypes, A1a, A1b, A2a, and A2b, as well as type B. Genotypic and geographic divisions observed among isolates from humans were mirrored among isolates from animals, specifically among animal species that are linked to human infection and to enzootic maintenance of tularemia. Significant differences between human infections caused by different genotypes were identified with respect to patient age, site of organism recovery, and mortality. Human infections due to A1b resulted in significantly higher mortality (24%) than those caused by A1a (4%), A2 (0%), and type B (7%). CONCLUSIONS: Three type A genotypes, A1a, A1b, and A2, were shown to be epidemiologically important. Our analysis suggests that A1b strains may be significantly more virulent in humans than A1a, A2, or type B strains. These findings have important implications for disease progression, disease prevention, and basic research programs.
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Francisella tularensis/classificação , Francisella tularensis/isolamento & purificação , Tularemia/microbiologia , Tularemia/veterinária , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Animais , Técnicas de Tipagem Bacteriana , Criança , Pré-Escolar , Análise por Conglomerados , Impressões Digitais de DNA , DNA Bacteriano/genética , Eletroforese em Gel de Campo Pulsado , Feminino , Francisella tularensis/genética , Francisella tularensis/patogenicidade , Genótipo , Humanos , Lactente , Masculino , Pessoa de Meia-Idade , Epidemiologia Molecular , Tularemia/epidemiologia , Tularemia/mortalidade , Estados Unidos/epidemiologia , Adulto JovemRESUMO
PulseNet, the National Molecular Subtyping Network for Foodborne Disease Surveillance, was established in 1996 through a collaboration with the Centers for Disease Control and Prevention; the US Department of Agriculture, Food Safety and Inspection Service; the US Food and Drug Administration; 4 state public health laboratories; and the Association of Public Health Laboratories. The network has since expanded to include 83 state, local, and food regulatory public health laboratories. In 2016, PulseNet was estimated to be helping prevent an estimated 270 000 foodborne illnesses annually. PulseNet is undergoing a transformation toward whole-genome sequencing (WGS), which provides better discriminatory power and precision than pulsed-field gel electrophoresis (PFGE). WGS improves the detection of outbreak clusters and could replace many traditional reference identification and characterization methods. This article highlights the contributions made by public health laboratories in transforming PulseNet's surveillance and describes how the transformation is changing local and national surveillance practices. Our data show that WGS is better at identifying clusters than PFGE, especially for clonal organisms such as Salmonella Enteritidis. The need to develop prioritization schemes for cluster follow-up and additional resources for both public health laboratory and epidemiology departments will be critical as PulseNet implements WGS for foodborne disease surveillance in the United States.
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Surtos de Doenças/prevenção & controle , Doenças Transmitidas por Alimentos/epidemiologia , Laboratórios , Vigilância em Saúde Pública , Saúde Pública , Eletroforese em Gel de Campo Pulsado , Humanos , Estados Unidos/epidemiologia , Sequenciamento Completo do GenomaRESUMO
We present TaqMan-minor groove binding (MGB) assays for an SNP that separates the Yersinia pestis strain CO92 from all other strains and for another SNP that separates North American strains from all other global strains.
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Técnicas de Tipagem Bacteriana/métodos , Yersinia pestis/classificação , Yersinia pestis/isolamento & purificação , Alelos , DNA Bacteriano/análise , DNA Bacteriano/genética , Genótipo , América do Norte , Polimorfismo de Nucleotídeo Único/genética , Reprodutibilidade dos Testes , Fatores de Tempo , Yersinia pestis/genéticaRESUMO
PulseNet USA is the national molecular subtyping network system for foodborne disease surveillance. Sixty-four public health and food regulatory laboratories participate in PulseNet USA and routinely perform pulsed-field gel electrophoresis of Shiga toxigenic Escherichia coli isolated from humans, food, water, and the environment on a real-time basis. Clusters of infection are detected in three ways within this system: through rapidly alerting the participants in the electronic communication forum, the PulseNet Web conference; through cluster analysis by the database administrators at the coordinating center at the Centers for Disease Control and Prevention of the patterns uploaded to the central server by the participants; and by matching profiles of strains from nonhuman sources with recent human uploads to the national server. The strengths, limitations, and scope for future improvements of PulseNet are discussed with examples from 2002. In that year, notices of 30 clusters of Shiga toxigenic E. coli O157 infections were posted on the Web conference, 26 of which represented local outbreaks, whereas four were multistate outbreaks. Another 27 clusters were detected by central cluster detection performed at the Centers for Disease Control and Prevention, of which five represented common source outbreaks confirmed after finding an isolate with the outbreak pattern in the implicated food. Ten food isolates submitted without suspicion of an association to human disease matched human isolates in the database, and an epidemiologic link to human cases was established for six of them.
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Técnicas de Tipagem Bacteriana , Eletroforese em Gel de Campo Pulsado/métodos , Escherichia coli O157/isolamento & purificação , Serviços de Informação , Saúde Pública , Análise por Conglomerados , Bases de Dados como Assunto , Surtos de Doenças , Escherichia coli O157/classificação , Microbiologia de Alimentos , Vigilância da População , Controle de Qualidade , Estados UnidosRESUMO
In August 2000, the Ohio Department of Health reported a cluster of men with typhoid fever who denied having traveled abroad. To determine the cause and the extent of the outbreak, an epidemiological investigation was initiated in which 7 persons in Ohio, Kentucky, and Indiana with culture-confirmed Salmonella enterica serotype Typhi infection and 2 persons with probable typhoid fever were evaluated; all were men, and all but one reported having had sex with 1 asymptomatic male S. Typhi carrier. We document sexual transmission of typhoid fever, which may be acquired by means of oral and anal sex, as well as via food and drink.
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Surtos de Doenças , Homossexualidade Masculina , Salmonella typhi , Infecções Sexualmente Transmissíveis/epidemiologia , Febre Tifoide/epidemiologia , Humanos , Indiana/epidemiologia , Kentucky/epidemiologia , Masculino , Ohio/epidemiologiaRESUMO
In December 2000, physicians in the Republic of the Marshall Islands reported the first known outbreak of Vibrio cholerae O1 infection (biotype El Tor, serotype Ogawa) from this country. In a matched case-control study on Ebeye Island, patients with cholera (n=53) had greater odds than persons without cholera (n=104) to have drunk adequately chlorinated water collected from a US military installation on neighboring Kwajalein Island and transported back to Ebeye (matched odds ratio [MOR], 8.0; P=.01). Transporting or storing drinking water in a water cooler with a spout and a tight-fitting lid was associated with reduced odds of illness (MOR, 0.24; P<.01), as was drinking bottled water (MOR, 0.08; P<.01), boiled water (MOR, 0.47; P=.02), or water flavored with powdered drink mixes (MOR, 0.18; P<.01). No cases of cholera were reported among Kwajalein residents. This outbreak highlights the critical importance of handling and storing drinking water safely, especially during outbreaks of gastrointestinal illness.
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Cólera/epidemiologia , Surtos de Doenças , Vibrio cholerae , Abastecimento de Água , Adolescente , Adulto , Idoso , Criança , Pré-Escolar , Cloro/farmacologia , Feminino , Humanos , Hidroterapia , Masculino , Micronésia/epidemiologia , Pessoa de Meia-Idade , Vibrio cholerae/efeitos dos fármacosRESUMO
Francisella tularensis is the causative agent of tularemia, which is a highly lethal disease from nature and potentially from a biological weapon. This species contains four recognized subspecies including the North American endemic F. tularensis subsp. tularensis (type A), whose genetic diversity is correlated with its geographic distribution including a major population subdivision referred to as A.I and A.II. The biological significance of the A.I - A.II genetic differentiation is unknown, though there are suggestive ecological and epidemiological correlations. In order to understand the differentiation at the genomic level, we have determined the complete sequence of an A.II strain (WY96-3418) and compared it to the genome of Schu S4 from the A.I population. We find that this A.II genome is 1,898,476 bp in size with 1,820 genes, 1,303 of which code for proteins. While extensive genomic variation exists between "WY96" and Schu S4, there is only one whole gene difference. This one gene difference is a hypothetical protein of unknown function. In contrast, there are numerous SNPs (3,367), small indels (1,015), IS element differences (7) and large chromosomal rearrangements (31), including both inversions and translocations. The rearrangement borders are frequently associated with IS elements, which would facilitate intragenomic recombination events. The pathogenicity island duplicated regions (DR1 and DR2) are essentially identical in WY96 but vary relative to Schu S4 at 60 nucleotide positions. Other potential virulence-associated genes (231) varied at 559 nucleotide positions, including 357 non-synonymous changes. Molecular clock estimates for the divergence time between A.I and A.II genomes for different chromosomal regions ranged from 866 to 2131 years before present. This paper is the first complete genomic characterization of a member of the A.II clade of Francisella tularensis subsp. tularensis.
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DNA Circular/genética , Francisella tularensis/genética , Genoma Bacteriano , Elementos de DNA Transponíveis/genética , DNA Bacteriano/química , DNA Bacteriano/genética , Francisella tularensis/isolamento & purificação , Francisella tularensis/patogenicidade , Ordem dos Genes , Humanos , Reação em Cadeia da Polimerase , Polimorfismo de Nucleotídeo Único , Análise de Sequência de DNA , Especificidade da Espécie , Virulência/genéticaRESUMO
Tularemia in the United States is caused by 2 subspecies of Francisella tularensis, subspecies tularensis (type A) and subspecies holarctica (type B). We compared clinical and demographic features of human tularemia cases from 1964 to 2004 from 39 states in which an isolate was recovered and subtyped. Our data indicate that type A and type B infections differ with respect to affected populations, anatomic site of isolation, and geographic distribution. Molecular subtyping with pulsed-field gel electrophoresis further defined 2 subpopulations of type A (type A-east and type A-west) that differ with respect to geographic distribution, disease outcome, and transmission. Our data suggest that type A-west infections are less severe than either type B or type A-east infections. Through a combined epidemiologic and molecular approach to human cases of tularemia, we provide new insights into the disease for future investigation.
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Tularemia/epidemiologia , Tularemia/microbiologia , Adulto , Criança , Feminino , Francisella tularensis/genética , Francisella tularensis/patogenicidade , Humanos , Masculino , Pessoa de Meia-Idade , Filogenia , Estações do Ano , Fatores de Tempo , Estados Unidos/epidemiologiaRESUMO
The causative agent of tularemia, Francisella tularensis, is a formidable biologic agent that occurs naturally throughout North America. We examined genetic and spatial diversity patterns among 161 US F. tularensis isolates by using a 24-marker multiple-locus variable-number tandem repeat analysis (MLVA) system. MLVA identified 126 unique genotypes. Phylogenetic analyses showed patterns similar to recently reported global-scale analyses. We observed clustering by subspecies, low genetic diversity within F. tularensis subsp. holarctica, and division of F. tularensis subsp. tularensis into 2 distinct subpopulations: A.I. and A.II. The 2 F. tularensis subsp. tularensis subpopulations also represent geographically distinct groups; A.I. occurs primarily in the central United States, and A.II. occurs primarily in the western United States. These spatial distributions are correlated with geographic ranges of particular vectors, hosts of tularemia, and abiotic factors. These correlates provide testable hypotheses regarding ecologic factors associated with maintaining tularemia foci.
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Francisella tularensis/genética , Francisella tularensis/isolamento & purificação , Tularemia/epidemiologia , Tularemia/microbiologia , Animais , Francisella tularensis/classificação , Geografia , Filogenia , Estados Unidos/epidemiologiaRESUMO
The PulseNet National Database, established by the Centers for Disease Control and Prevention in 1996, consists of pulsed-field gel electrophoresis (PFGE) patterns obtained from isolates of food-borne pathogens (currently Escherichia coli O157:H7, Salmonella, Shigella, and Listeria) and textual information about the isolates. Electronic images and accompanying text are submitted from over 60 U.S. public health and food regulatory agency laboratories. The PFGE patterns are generated according to highly standardized PFGE protocols. Normalization and accurate comparison of gel images require the use of a well-characterized size standard in at least three lanes of each gel. Originally, a well-characterized strain of each organism was chosen as the reference standard for that particular database. The increasing number of databases, difficulty in identifying an organism-specific standard for each database, the increased range of band sizes generated by the use of additional restriction endonucleases, and the maintenance of many different organism-specific strains encouraged us to search for a more versatile and universal DNA size marker. A Salmonella serotype Braenderup strain (H9812) was chosen as the universal size standard. This strain was subjected to rigorous testing in our laboratories to ensure that it met the desired criteria, including coverage of a wide range of DNA fragment sizes, even distribution of bands, and stability of the PFGE pattern. The strategy used to convert and compare data generated by the new and old reference standards is described.
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Bases de Dados como Assunto , Eletroforese em Gel de Campo Pulsado/normas , Eletroforese em Gel de Campo Pulsado/métodos , Escherichia coli O157/genética , Listeria monocytogenes/genética , Padrões de Referência , Salmonella/genética , SorotipagemRESUMO
Toxigenic Vibrio cholerae serogroup O141 has been associated with sporadic cholera-like diarrhea and bloodstream infection in the United States. Consumption of seafood and proximity to the coast may increase the risk of infection. All V. cholerae isolates recovered from stool samples of patients with diarrhea or from a normally sterile site should be serogrouped and assessed for cholera toxin production. Improved surveillance and case-control studies are needed to further characterize illness and risk factors for V. cholerae O141 infection.