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1.
Anal Chem ; 96(17): 6588-6598, 2024 Apr 30.
Artigo em Inglês | MEDLINE | ID: mdl-38619494

RESUMO

How timely identification and determination of pathogen species in pathogen-contaminated foods are responsible for rapid and accurate treatments for food safety accidents. Herein, we synthesize four aggregation-induced emissive nanosilicons with different surface potentials and hydrophobicities by encapsulating four tetraphenylethylene derivatives differing in functional groups. The prepared nanosilicons are utilized as receptors to develop a nanosensor array according to their distinctive interactions with pathogens for the rapid and simultaneous discrimination of pathogens. By coupling with machine-learning algorithms, the proposed nanosensor array achieves high performance in identifying eight pathogens within 1 h with high overall accuracy (93.75-100%). Meanwhile, Cronobacter sakazakii and Listeria monocytogenes are taken as model bacteria for the quantitative evaluation of the developed nanosensor array, which can successfully distinguish the concentration of C. sakazakii and L. monocytogenes at more than 103 and 102 CFU mL-1, respectively, and their mixed samples at 105 CFU mL-1 through the artificial neural network. Moreover, eight pathogens at 1 × 104 CFU mL-1 in milk can be successfully identified by the developed nanosensor array, indicating its feasibility in monitoring food hazards.


Assuntos
Microbiologia de Alimentos , Listeria monocytogenes , Aprendizado de Máquina , Listeria monocytogenes/isolamento & purificação , Cronobacter sakazakii/isolamento & purificação , Dióxido de Silício/química , Sistemas Automatizados de Assistência Junto ao Leito , Animais , Leite/microbiologia , Leite/química , Técnicas Biossensoriais , Redes Neurais de Computação
2.
Anal Chem ; 96(21): 8543-8551, 2024 May 28.
Artigo em Inglês | MEDLINE | ID: mdl-38748432

RESUMO

In this study, the covalently fixed "end-on" orientation of a monoclonal Listeria monocytogenes antibody (mAb-Lis) to amino terminated oligo (ethylene glycol)-capped gold nanoparticles (NH2-TEG-AuNPs) was used to fabricate an in-house lateral flow strip (LFS), namely, the fixed "end-on" Lis-mAb-NH-TEG-AuNPs LFS. The aim was to evaluate the performance of the fixed "end-on" Lis-mAb-NH-TEG-AuNPs LFS in detecting L. monocytogenes. The proposed LFS enabled the sensitive detection of L. monocytogenes in 15 min with a visual limit of detection of 102 CFU/mL. Quantitative analysis indicated an LOD at 10 CFU/mL. The fixed "end-on" Lis-mAb-NH-TEG-AuNPs LFS showed no cross-reactivity with other pathogenic bacteria and practical performance across different food matrices, including human blood, milk, and mushroom samples. Furthermore, the clinical performance of the fixed "end-on" Lis-mAb-NH-TEG-AuNPs LFS for detecting L. monocytogenes was evaluated by using 12 clinical samples validated by the hemoculture method. It demonstrated excellent concordance with the reference methods, with no false-positive or false-negative results observed. Therefore, the fixed "end-on" Lis-mAb-NH-TEG-AuNPs LFS serves as a promising candidate for a point-of-care test (POCT), enabling the rapid, precise, and highly sensitive detection of L. monocytogenes in clinical samples and contaminated food.


Assuntos
Anticorpos Monoclonais , Ouro , Listeria monocytogenes , Nanopartículas Metálicas , Listeria monocytogenes/isolamento & purificação , Listeria monocytogenes/imunologia , Ouro/química , Nanopartículas Metálicas/química , Anticorpos Monoclonais/imunologia , Anticorpos Monoclonais/química , Humanos , Limite de Detecção , Microbiologia de Alimentos , Leite/microbiologia , Leite/química , Anticorpos Antibacterianos/química , Anticorpos Antibacterianos/imunologia , Anticorpos Imobilizados/química , Anticorpos Imobilizados/imunologia , Animais , Listeriose/microbiologia , Listeriose/diagnóstico
3.
J Clin Microbiol ; 62(5): e0157623, 2024 May 08.
Artigo em Inglês | MEDLINE | ID: mdl-38441926

RESUMO

Whole-genome sequencing has become the method of choice for bacterial outbreak investigation, with most clinical and public health laboratories currently routinely using short-read Illumina sequencing. Recently, long-read Oxford Nanopore Technologies (ONT) sequencing has gained prominence and may offer advantages over short-read sequencing, particularly with the recent introduction of the R10 chemistry, which promises much lower error rates than the R9 chemistry. However, limited information is available on its performance for bacterial single-nucleotide polymorphism (SNP)-based outbreak investigation. We present an open-source workflow, Prokaryotic Awesome variant Calling Utility (PACU) (https://github.com/BioinformaticsPlatformWIV-ISP/PACU), for constructing SNP phylogenies using Illumina and/or ONT R9/R10 sequencing data. The workflow was evaluated using outbreak data sets of Shiga toxin-producing Escherichia coli and Listeria monocytogenes by comparing ONT R9 and R10 with Illumina data. The performance of each sequencing technology was evaluated not only separately but also by integrating samples sequenced by different technologies/chemistries into the same phylogenomic analysis. Additionally, the minimum sequencing time required to obtain accurate phylogenetic results using nanopore sequencing was evaluated. PACU allowed accurate identification of outbreak clusters for both species using all technologies/chemistries, but ONT R9 results deviated slightly more from the Illumina results. ONT R10 results showed trends very similar to Illumina, and we found that integrating data sets sequenced by either Illumina or ONT R10 for different isolates into the same analysis produced stable and highly accurate phylogenomic results. The resulting phylogenies for these two outbreaks stabilized after ~20 hours of sequencing for ONT R9 and ~8 hours for ONT R10. This study provides a proof of concept for using ONT R10, either in isolation or in combination with Illumina, for rapid and accurate bacterial SNP-based outbreak investigation.


Assuntos
Surtos de Doenças , Polimorfismo de Nucleotídeo Único , Humanos , Sequenciamento por Nanoporos/métodos , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Filogenia , Listeria monocytogenes/genética , Listeria monocytogenes/classificação , Listeria monocytogenes/isolamento & purificação , Sequenciamento Completo do Genoma/métodos , Genoma Bacteriano/genética , Listeriose/epidemiologia , Listeriose/microbiologia , Análise de Sequência de DNA/métodos , Nanoporos , Bactérias/genética , Bactérias/classificação , Bactérias/isolamento & purificação
4.
BMC Infect Dis ; 24(1): 477, 2024 May 08.
Artigo em Inglês | MEDLINE | ID: mdl-38720244

RESUMO

We report a very rare case of Listeria multiple brain abscesses manifested as delirium, which represented diagnostic and therapeutic challenges overcome only by the close cooperation between Infectious Diseases and Neuroradiology, without which a satisfactory outcome would not be achieved.An elderly man presented with confusion and drowsiness with a background of type-II diabetes mellitus. Although computed tomography of the brain only showed frontal lobe oedema, contrast magnetic resonance (MR) imaging showed numerous irregular rim-enhancing lesions containing central diffusion restriction, suggesting multiple pyogenic cerebral abscesses of unclear aetiology. Thereafter, Listeria monocytogenes was isolated from blood cultures, suggesting this as the causative organism. Deemed unsuitable for neurosurgical drainage, the patient received medical management with a protracted course of antibiotics. This case was extremely challenging, due to 1) the impossibility of source control, 2) the small number of effective antibiotics available to treat this condition, and 3) the inevitable antibiotic side-effects, derived from long-term exposure. A successful outcome was only possible thanks to strict close multidisciplinary follow up, requiring frequent MR imaging and a judicious antibiotic choice, including monitoring of their side-effects. Due to the rarity of this condition, there is lack of guidance on its management, hence the importance of multidisciplinary involvement with very close imaging and antibiotic monitoring.


Assuntos
Antibacterianos , Abscesso Encefálico , Listeria monocytogenes , Listeriose , Humanos , Masculino , Abscesso Encefálico/microbiologia , Abscesso Encefálico/tratamento farmacológico , Abscesso Encefálico/diagnóstico por imagem , Listeriose/tratamento farmacológico , Listeriose/microbiologia , Listeriose/diagnóstico , Antibacterianos/uso terapêutico , Listeria monocytogenes/isolamento & purificação , Idoso , Imageamento por Ressonância Magnética , Tomografia Computadorizada por Raios X , Encéfalo/diagnóstico por imagem , Encéfalo/patologia , Encéfalo/microbiologia , Delírio/tratamento farmacológico
5.
BMC Vet Res ; 20(1): 297, 2024 Jul 06.
Artigo em Inglês | MEDLINE | ID: mdl-38971767

RESUMO

BACKGROUND: Listeriosis is a global health threat to both animals and humans, especially in developing countries. This study was designed to isolate Listeria monocytogenes from faeces; environmental samples; and cow, sheep and goat milk, as well as human stool, to study its molecular characteristics and antibiotic sensitivity in the New Valley and Beheira Governorates, Egypt. The isolation and identification of L. monocytogenes were carried out using traditional culture and biochemical methods, followed by antibiography, genus confirmation of some isolates and detection and sequencing of InlB genes via PCR. RESULTS: Out of 2097 examined samples, the prevalence of L. monocytogenes was 13.4% in animals; the prevalence was 9.2%, 2.4%, 25.4%, 4%, 42.4%, and 6.4% in cattle faeces, cattle milk, sheep faeces, sheep milk, goat faeces, and goat milk, respectively. However, the prevalence of L. monocytogenes was 8.3% in human samples. Both animal and human isolates showed 100% resistance to trimethoprim-sulfamethoxazole, and the isolates showed the highest sensitivity to flumequine (100%), amikacin (99.2%), gentamicin (97.6%), and levofloxacin (94.6%). Multidrug resistance (MDR) was detected in 86.9% of the tested isolates. The 16 S rRNA and inlB genes were detected in 100% of the randomly selected L. monocytogenes isolates. Phylogenetic analysis of three isolates based on the inlB gene showed 100% identity between faecal, milk and human stool isolates. CONCLUSIONS: Faeces and milk are major sources of listeriosis, and the high degree of genetic similarity between animal and human isolates suggests the possibility of zoonotic circulation. The high prevalence of MDR L. monocytogenes in both animal and human samples could negatively impact the success of prevention and treatments for animal and human diseases, thereby imposing serious risks to public health.


Assuntos
Antibacterianos , Fezes , Cabras , Listeria monocytogenes , Listeriose , Leite , Animais , Egito/epidemiologia , Listeria monocytogenes/efeitos dos fármacos , Listeria monocytogenes/genética , Listeria monocytogenes/isolamento & purificação , Humanos , Prevalência , Ovinos , Antibacterianos/farmacologia , Bovinos , Fezes/microbiologia , Listeriose/veterinária , Listeriose/epidemiologia , Listeriose/microbiologia , Leite/microbiologia , Testes de Sensibilidade Microbiana , Farmacorresistência Bacteriana/genética
6.
Euro Surveill ; 29(19)2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-38726694

RESUMO

Listeria monocytogenes (Lm) is a bacterium widely distributed in the environment. Listeriosis is a severe disease associated with high hospitalisation and mortality rates. In April 2019, listeriosis was diagnosed in two hospital patients in Finland. We conducted a descriptive study to identify the source of the infection and defined a case as a person with a laboratory-confirmed Lm serogroup IIa sequence type (ST) 37. Six cases with Lm ST 37 were notified to the Finnish Infectious Diseases Registry between 2015 and 2019. Patient interviews and hospital menus were used to target traceback investigation of the implicated foods. In 2021 and 2022, similar Lm ST 37 was detected from samples of a ready-to-eat plant-based food product including fava beans. Inspections by the manufacturer and the local food control authority indicated that the food products were contaminated with Lm after pasteurisation. Our investigation highlights the importance that companies producing plant-based food are subject to similar controls as those producing food of animal origin. Hospital menus can be a useful source of information that is not dependent on patient recall.


Assuntos
Surtos de Doenças , Microbiologia de Alimentos , Listeria monocytogenes , Listeriose , Humanos , Listeria monocytogenes/isolamento & purificação , Listeria monocytogenes/genética , Listeriose/epidemiologia , Listeriose/microbiologia , Finlândia/epidemiologia , Feminino , Masculino , Doenças Transmitidas por Alimentos/epidemiologia , Doenças Transmitidas por Alimentos/microbiologia , Pessoa de Meia-Idade , Idoso , Contaminação de Alimentos , Adulto , Fabaceae/microbiologia
7.
Foodborne Pathog Dis ; 21(6): 386-394, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38346310

RESUMO

Listeria monocytogenes is a critical foodborne pathogen that causes severe invasive and noninvasive diseases and is associated with high mortality. Information on the prevalence of L. monocytogenes infections in Taiwan is very limited. This study aimed to analyze the molecular epidemiological surveillance and virulence gene distribution of 176 human clinical L. monocytogenes isolates collected between 2009 and 2019 in northern Taiwan. Our results showed that the isolates belonged to 4 serogroups (IIa, IIb, IVb, and IIc), with most isolates in serogroups IIa (81/176, 46%) and IIb (71/176, 40.3%). Multilocus sequence typing analysis revealed 18 sequence types (STs) and 13 clonal complexes (CCs). Eighty-four percent of all isolates belonged to six STs: CC87-ST87 (40/176, 22.7%), CC19-ST378 (36/176, 19.9%), CC155-ST155 (28/176, 15.5%), CC1-ST710 (16/176, 8.8%), CC5-ST5 (16/176, 8.8%), and CC101-ST101 (11/176, 6.1%). Furthermore, our analysis showed the distributions of four Listeria pathogenicity islands (LIPI) among all isolates. LIPI-1 and LIPI-2 existed in all isolates, whereas LIPI-3 and LIPI-4 only existed in specific STs and CCs. LIPI-3 existed in the STs, CC1-ST710, CC3-ST3, CC288-ST295, and CC191-ST1458, whereas LIPI-4 could be found in the STs, CC87-ST87 and CC87-ST1459. Strains containing LIPI-3 and LIPI-4 are potentially hypervirulent; thus, 68/176 isolates (39.1%) collected in this study were potentially hypervirulent. Since L. monocytogenes infections are considered highly correlated with diet, molecular epidemiological surveillance of Listeria in food is important; continued surveillance will provide critical information to prevent foodborne diseases.


Assuntos
Listeria monocytogenes , Listeriose , Tipagem de Sequências Multilocus , Listeria monocytogenes/genética , Listeria monocytogenes/patogenicidade , Listeria monocytogenes/isolamento & purificação , Listeria monocytogenes/classificação , Taiwan/epidemiologia , Humanos , Listeriose/microbiologia , Listeriose/epidemiologia , Virulência/genética , Sorogrupo , Fatores de Virulência/genética , Ilhas Genômicas , Doenças Transmitidas por Alimentos/microbiologia , Doenças Transmitidas por Alimentos/epidemiologia , Epidemiologia Molecular
8.
Mikrochim Acta ; 191(7): 429, 2024 06 28.
Artigo em Inglês | MEDLINE | ID: mdl-38942915

RESUMO

The engineering of a home-made portable double-layer filtration and concentration device with the common syringe for rapid analysis of water samples is reported. The core elements of the device were two installed filtration membranes with different pore sizes for respective functions. The upper filtration membrane was used for preliminary intercepting large interfering impurities (interception membrane), while the lower filtration membrane was used for collecting multiple target pathogens (enrichment membrane) for determination. This combination can make the contaminated environmental water, exemplified by surface water, filtrated quickly through the device and just retained the target bacteria of Escherichia coli O157:H7, Staphylococcus aureus, and Listeria monocytogenes on the lower enrichment membrane. Integrating with surface-enhanced Raman spectra (SERS) platform to decode the SERS-Tags (SERS-TagCVa, SERS-TagR6G, and SERS-TagMB) already labeled on each of the enriched bacteria based the antibody-mediated immuno-recognition effect, fast separation, concentration, and detection of multiple pathogenic bacteria from the bulk of contaminated environmental water were realized. Results show that within 30 min, all target bacteria in the lake water can be simultaneously and accurately measured in the range from 101 to 106 CFU mL-1 with detection limit of 10.0 CFU mL-1 without any pre-culture procedures. This work highlights the simplicity, rapidness, cheapness, selectivity, and the robustness of the constructed method for simultaneous detecting multiple pathogens in aqueous samples. This protocol opens a new avenue for facilitating the development of versatile analytical tools for drinking water and food safety monitoring in underdeveloped or developing countries.


Assuntos
Água Potável , Escherichia coli O157 , Filtração , Limite de Detecção , Listeria monocytogenes , Análise Espectral Raman , Staphylococcus aureus , Análise Espectral Raman/métodos , Água Potável/microbiologia , Filtração/instrumentação , Staphylococcus aureus/isolamento & purificação , Listeria monocytogenes/isolamento & purificação , Escherichia coli O157/isolamento & purificação , Nanopartículas Metálicas/química , Microbiologia da Água
9.
Mikrochim Acta ; 191(6): 361, 2024 06 01.
Artigo em Inglês | MEDLINE | ID: mdl-38822891

RESUMO

A one-shot CO2 laser-based strategy to generate conductive reduced graphene oxide (rGO) decorated with nanoceria (nCe) is proposed. The 2D/0D rGO-nCe films, integrated as catalytic sensing layers in paper-based sensors, were employed for on-site monitoring of indoor fogging treatments against Listeria monocytogenes (Lm), a ubiquitous pathogenic bacterium. The rGO-nCe laser-assisted synthesis was optimized to preserve the rGO film morphological and electron-transfer features and simultaneously integrate catalytic nCe. The films were characterized by microscopical (SEM), spectroscopical (EDX, Raman, and FTIR), and electrochemical techniques. The most performing film was integrated into a nitrocellulose substrate, and the complete sensor was assembled via a combination of xurography and stencil printing. The rGO-nCe sensor's catalytic activity was proved toward the detection of H2O2, obtaining sensitive determination (LOD = 0.3 µM) and an extended linear range (0.5-1500 µM). Eventually, the rGO-nCe sensor was challenged for the real-time continuous monitoring of hydrogen peroxide aerosol during no-touch fogging treatment conducted following the EU's recommendation for biocidal product use. Treatment effectiveness was proved toward three Lm strains characterized by different origins, i.e., type strain ATCC 7644, clinical strain 338, and food strain 641/6II. The sensor allows for discrimination and quantification treatments at different environmental biocidal amounts and fogging times, and correlates with the microbiological inhibition, promoting the proposed sensor as a useful tool to modulate and monitor no-touch treatments.


Assuntos
Desinfecção , Grafite , Peróxido de Hidrogênio , Lasers , Listeria monocytogenes , Papel , Grafite/química , Peróxido de Hidrogênio/química , Listeria monocytogenes/efeitos dos fármacos , Listeria monocytogenes/isolamento & purificação , Desinfecção/métodos , Cério/química , Limite de Detecção , Técnicas Eletroquímicas/métodos , Técnicas Eletroquímicas/instrumentação , Catálise
10.
Sensors (Basel) ; 24(8)2024 Apr 19.
Artigo em Inglês | MEDLINE | ID: mdl-38676242

RESUMO

Foodborne pathogens are microbes present in food that cause serious illness when the contaminated food is consumed. Among these pathogens, Listeria monocytogenes is one of the most serious bacterial pathogens, and causes severe illness. The techniques currently used for L. monocytogenes detection are based on common molecular biology tools that are not easy to implement for field use in food production and distribution facilities. This work focuses on the efficacy of an electrochemical biosensor in detecting L. monocytogenes in chicken broth. The sensor is based on a nanostructured electrode modified with a bacteriophage as a bioreceptor which selectively detects L. monocytogenes using electrochemical impedance spectroscopy. The biosensing platform was able to reach a limit of detection of 55 CFU/mL in 1× PBS buffer and 10 CFU/mL in 1% diluted chicken broth. The biosensor demonstrated 83-98% recovery rates in buffer and 87-96% in chicken broth.


Assuntos
Técnicas Biossensoriais , Galinhas , Espectroscopia Dielétrica , Microbiologia de Alimentos , Listeria monocytogenes , Listeria monocytogenes/isolamento & purificação , Técnicas Biossensoriais/métodos , Animais , Microbiologia de Alimentos/métodos , Eletrodos
11.
N Engl J Med ; 382(7): 632-643, 2020 02 13.
Artigo em Inglês | MEDLINE | ID: mdl-32053299

RESUMO

BACKGROUND: An outbreak of listeriosis was identified in South Africa in 2017. The source was unknown. METHODS: We conducted epidemiologic, trace-back, and environmental investigations and used whole-genome sequencing to type Listeria monocytogenes isolates. A case was defined as laboratory-confirmed L. monocytogenes infection during the period from June 11, 2017, to April 7, 2018. RESULTS: A total of 937 cases were identified, of which 465 (50%) were associated with pregnancy; 406 of the pregnancy-associated cases (87%) occurred in neonates. Of the 937 cases, 229 (24%) occurred in patients 15 to 49 years of age (excluding those who were pregnant). Among the patients in whom human immunodeficiency virus (HIV) status was known, 38% of those with pregnancy-associated cases (77 of 204) and 46% of the remaining patients (97 of 211) were infected with HIV. Among 728 patients with a known outcome, 193 (27%) died. Clinical isolates from 609 patients were sequenced, and 567 (93%) were identified as sequence type 6 (ST6). In a case-control analysis, patients with ST6 infections were more likely to have eaten polony (a ready-to-eat processed meat) than those with non-ST6 infections (odds ratio, 8.55; 95% confidence interval, 1.66 to 43.35). Polony and environmental samples also yielded ST6 isolates, which, together with the isolates from the patients, belonged to the same core-genome multilocus sequence typing cluster with no more than 4 allelic differences; these findings showed that polony produced at a single facility was the outbreak source. A recall of ready-to-eat processed meat products from this facility was associated with a rapid decline in the incidence of L. monocytogenes ST6 infections. CONCLUSIONS: This investigation showed that in a middle-income country with a high prevalence of HIV infection, L. monocytogenes caused disproportionate illness among pregnant girls and women and HIV-infected persons. Whole-genome sequencing facilitated the detection of the outbreak and guided the trace-back investigations that led to the identification of the source.


Assuntos
Surtos de Doenças , Doenças Transmitidas por Alimentos/epidemiologia , Listeria monocytogenes/isolamento & purificação , Listeriose/epidemiologia , Produtos da Carne/microbiologia , Adolescente , Adulto , Idoso , Técnicas de Tipagem Bacteriana , Estudos de Casos e Controles , Feminino , Doenças Transmitidas por Alimentos/etiologia , Doenças Transmitidas por Alimentos/mortalidade , Infecções por HIV/complicações , HIV-1 , Humanos , Recém-Nascido , Listeria monocytogenes/genética , Listeriose/etiologia , Listeriose/mortalidade , Masculino , Produtos da Carne/efeitos adversos , Pessoa de Meia-Idade , Gravidez , Complicações Infecciosas na Gravidez/epidemiologia , Recall e Retirada de Produto , Distribuição por Sexo , África do Sul/epidemiologia , Sequenciamento Completo do Genoma , Adulto Jovem
12.
Proc Natl Acad Sci U S A ; 117(22): 11923-11930, 2020 06 02.
Artigo em Inglês | MEDLINE | ID: mdl-32414933

RESUMO

Here we report a sensing method for Listeria monocytogenes based on the agglutination of all-liquid Janus emulsions. This two-dye assay enables the rapid detection of trace Listeria in less than 2 h via an emissive signal produced in response to Listeria binding. The biorecognition interface between the Janus emulsions is assembled by attaching antibodies to a functional surfactant polymer with a tetrazine/transcyclooctene click reaction. The strong binding between Listeria and the Listeria antibody located at the hydrocarbon surface of the emulsions results in the tilting of the Janus structure from its equilibrium position to produce emission that would ordinarily be obscured by a blocking dye. This method provides rapid and inexpensive Listeria detection with high sensitivity (<100 CFU/mL in 2 h) that can be paired with many antibody or related recognition elements to create a new class of biosensors.


Assuntos
Técnicas Biossensoriais/métodos , Emulsões/química , Corantes Fluorescentes/química , Listeria monocytogenes/isolamento & purificação , Aglutinação , Anticorpos Antibacterianos , Contaminação de Alimentos/análise , Microbiologia de Alimentos/métodos , Doenças Transmitidas por Alimentos/prevenção & controle , Listeria monocytogenes/imunologia , Listeriose/prevenção & controle , Microscopia Confocal
13.
J Dairy Sci ; 105(12): 9450-9462, 2022 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-36207178

RESUMO

Foodborne pathogens detection is important to ensure food safety and human health. In this study, we designed a comet structure to rapidly and sensitively detect foodborne Listeria monocytogenes. This method combined isothermal sequence exchange amplification (SEA) and surface-enhanced Raman spectroscopy. Listeria monocytogenes DNA could be rapidly amplified at a constant temperature via SEA with a pair of modified primers, which rendered the precise thermal control instrumentation unnecessary. Efficient SEA amplification generated a large number of DNA duplexes that could be easily captured by streptavidin-modified magnetic bead and AuMB@Ag-isothiocyanate fluorescein antibody (anti-FITC). AuMB@Ag-anti-FITC was used as a signal probe, which generated a significant excitation signal at 1,616 cm-1 for quantitative detection and analysis. The results displayed sensitive detection of L. monocytogenes in cheese from 2.0 × 101 cfu/mL to 2.0 × 106 cfu/mL within 1.0 h with a detection limit of 7.8 cfu/mL. Furthermore, this comet structure displayed the desirable specificity as its specific primers and amplified DNA ends were attached to streptavidin-modified magnetic beads and AuMB@Ag-anti-FITC, respectively. We expected that the method devised would provide a promising new approach to screening for L. monocytogenes and guarantee the microbiological safety of dairy products.


Assuntos
Queijo , Contaminação de Alimentos , Listeria monocytogenes , Queijo/microbiologia , Primers do DNA/genética , Microbiologia de Alimentos , Listeria monocytogenes/isolamento & purificação , Análise Espectral Raman , Estreptavidina
14.
Mikrochim Acta ; 189(3): 94, 2022 02 08.
Artigo em Inglês | MEDLINE | ID: mdl-35132460

RESUMO

Recent research in the field of electrochemical biosensors equipped with peptides and nanomaterials have been categorized, reviewed, and critically analyzed. Indeed, using these innovative biosensors can revolutionize biomedical diagnostics in the future. Saving lives, time, and money in this field will be considered as some main benefits of this type of diagnosis. Here, these biosensors have been categorized and evaluated in four main sections. In the first section, the focus is on investigating the types of electrochemical peptide-based nanobiosensors applied to detect pathogenic microorganisms, microbial toxins, and viruses. In the second section, due to the importance of rapid diagnosis and prognosis of various cancers, the electrochemical peptide-based nanobiosensors designed to detect cancer biomarkers have been reviewed and analyzed. In the third section, the electrochemical peptide-based nanobiosensors, which were applied to detect the essential and effective biomolecules in the various diseases, and health control, including enzymes, hormones, biomarkers, and other biomolecules, have been considered. Finally, using a comprehensive analysis, all the used elements in these biosensors have been presented as conceptual diagrams that can effectively guide researchers in future developments. The essential factors in evaluating and analyzing these electrochemical peptide-based nanobiosensors such as analyte, peptide sequence, functional groups interacted between the peptide sequences and other biosensing components, the applied nanomaterials, diagnostic techniques, detection range, and limit of detection have also been included. Other analyzable items such as the type of used redox marker and the location of the peptide sequence against the signal transducer were also considered.


Assuntos
Técnicas Biossensoriais , Técnicas Eletroquímicas , Neoplasias/diagnóstico , Peptídeos/química , Humanos , Listeria monocytogenes/isolamento & purificação , Nanoestruturas/química , Proteínas/análise , Staphylococcus aureus/isolamento & purificação
15.
Appl Environ Microbiol ; 87(24): e0177421, 2021 11 24.
Artigo em Inglês | MEDLINE | ID: mdl-34613762

RESUMO

In this study, we addressed different aspects regarding the implementation of quasimetagenomic sequencing as a hybrid surveillance method in combination with enrichment for early detection of Listeria monocytogenes in the food industry. Different experimental enrichment cultures were used, comprising seven L. monocytogenes strains of different sequence types (STs), with and without a background microbiota community. To assess whether the proportions of the different STs changed over time during enrichment, the growth and population dynamics were assessed using dapE colony sequencing and dapE and 16S rRNA amplicon sequencing. There was a tendency of some STs to have a higher relative abundance during the late stage of enrichment when L. monocytogenes was enriched without background microbiota. When coenriched with background microbiota, the population dynamics of the different STs was more consistent over time. To evaluate the earliest possible time point during enrichment that allows the detection of L. monocytogenes and at the same time the generation of genetic information that enables an estimation regarding the strain diversity in a sample, quasimetagenomic sequencing was performed early during enrichment in the presence of the background microbiota using Oxford Nanopore Technologies Flongle and Illumina MiSeq sequencing. The application of multiple displacement amplification (MDA) enabled detection of L. monocytogenes (and the background microbiota) after only 4 h of enrichment using both applied sequencing approaches. The MiSeq sequencing data additionally enabled the prediction of cooccurring L. monocytogenes strains in the samples. IMPORTANCE We showed that a combination of a short primary enrichment combined with MDA and Nanopore sequencing can accelerate the traditional process of cultivation and identification of L. monocytogenes. The use of Illumina MiSeq sequencing additionally allowed us to predict the presence of cooccurring L. monocytogenes strains. Our results suggest quasimetagenomic sequencing is a valuable and promising hybrid surveillance tool for the food industry that enables faster identification of L. monocytogenes during early enrichment. Routine application of this approach could lead to more efficient and proactive actions in the food industry that prevent contamination and subsequent product recalls and food destruction, economic and reputational losses, and human listeriosis cases.


Assuntos
Microbiologia de Alimentos , Listeria monocytogenes , Microbiota , Genes Bacterianos , Listeria monocytogenes/genética , Listeria monocytogenes/isolamento & purificação , Metagenômica , Dinâmica Populacional , RNA Ribossômico 16S/genética
16.
Crit Rev Microbiol ; 47(5): 647-666, 2021 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-33896354

RESUMO

Listeria monocytogenes is one of the most invasive foodborne pathogens and is responsible for numerous outbreaks worldwide. Most of the methods to detect this bacterium in food require selective enrichment using traditional bacterial culture techniques that can be time-consuming and labour-intensive. Moreover, molecular methods are expensive and need specific technical knowledge. In contrast, immunological approaches are faster, simpler, and user-friendly alternatives and have been developed for the detection of L. monocytogenes in food, environmental, and clinical samples. These techniques are dependent on the constitutive expression of L. monocytogenes antigens and the specificity of the antibodies used. Here, updated knowledge on pathogenesis and the key immunogenic virulence determinants of L. monocytogenes that are used for the generation of monoclonal and polyclonal antibodies for the serological assay development are summarised. In addition, immunological approaches based on enzyme-linked immunosorbent assay, immunofluorescence, lateral flow immunochromatographic assays, and immunosensors with relevant improvements are highlighted. Though the sensitivity and specificity of the assays were improved significantly, methods still face many challenges that require further validation before use.


Assuntos
Listeria monocytogenes/isolamento & purificação , Listeria monocytogenes/patogenicidade , Listeriose/microbiologia , Fatores de Virulência/análise , Fatores de Virulência/imunologia , Animais , Anticorpos Antibacterianos/imunologia , Antígenos de Bactérias/análise , Antígenos de Bactérias/imunologia , Técnicas Biossensoriais , Microbiologia de Alimentos , Humanos , Imunidade Inata , Listeria monocytogenes/crescimento & desenvolvimento , Listeria monocytogenes/imunologia , Listeriose/diagnóstico , Listeriose/imunologia , Virulência , Fatores de Virulência/metabolismo
17.
PLoS Comput Biol ; 16(12): e1008468, 2020 12.
Artigo em Inglês | MEDLINE | ID: mdl-33378360

RESUMO

Multiplex polymerase chain reaction (PCR) has multiple applications in molecular biology, including developing new targeted next-generation sequencing (NGS) panels. We present NGS-PrimerPlex, an efficient and versatile command-line application that designs primers for different refined types of amplicon-based genome target enrichment. It supports nested and anchored multiplex PCR, redistribution among multiplex reactions of primers constructed earlier, and extension of existing NGS-panels. The primer design process takes into consideration the formation of secondary structures, non-target amplicons between all primers of a pool, primers and high-frequent genome single-nucleotide polymorphisms (SNPs) overlapping. Moreover, users of NGS-PrimerPlex are free from manually defining input genome regions, because it can be done automatically from a list of genes or their parts like exon or codon numbers. Using the program, the NGS-panel for sequencing the LRRK2 gene coding regions was created, and 354 DNA samples were studied successfully with a median coverage of 97.4% of target regions by at least 30 reads. To show that NGS-PrimerPlex can also be applied for bacterial genomes, we designed primers to detect foodborne pathogens Salmonella enterica, Escherichia coli O157:H7, Listeria monocytogenes, and Staphylococcus aureus considering variable positions of the genomes.


Assuntos
Sequenciamento de Nucleotídeos em Larga Escala/métodos , Reação em Cadeia da Polimerase Multiplex/métodos , Códon , Escherichia coli O157/genética , Escherichia coli O157/isolamento & purificação , Éxons , Genes Bacterianos , Genes Virais , Serina-Treonina Proteína Quinase-2 com Repetições Ricas em Leucina/genética , Listeria monocytogenes/genética , Listeria monocytogenes/isolamento & purificação , Hibridização de Ácido Nucleico , Polimorfismo de Nucleotídeo Único , Staphylococcus aureus/genética , Staphylococcus aureus/isolamento & purificação , Vírus/genética
18.
Nanotechnology ; 32(8): 085502, 2021 Feb 19.
Artigo em Inglês | MEDLINE | ID: mdl-33202394

RESUMO

The immunochromatographic strip test (ICST) is a powerful on-site detection technology due to its unique advantages of simplicity, rapidity, and readability by the naked eye. Here we illustrate the potential of α-Fe2O3 polyhedrons as a novel visual label, which exhibit advantages of high stability and economy, for the detection of Listeria monocytogenes (L. monocytogenes) as a model foodborne pathogen. A low-cost and simple one-step solvothermal approach was developed for the synthesis of α-Fe2O3 polyhedrons; the average diameter of the α-Fe2O3 polyhedrons is about 200 nm. The crystal structure and morphology of α-Fe2O3 polyhedrons were characterized by x-ray diffraction and transmission electron microscope. α-Fe2O3 polyhedrons were immunized with anti-L. monocytogenes antibody to prepare an antibody-colloidal α-Fe2O3 polyhedron ICST. Visual detection can be obtained directly by the naked eye within 10 min. The detection limit of L. monocytogenes by α-Fe2O3 polyhedron ICST assay was 3.8 × 106 and 5.6 × 106 CFU/ml of pure culture and artificially spiked orange juice drink sample, respectively. Results indicated that the antibody-colloidal α-Fe2O3 polyhedron ICST is a rapid, simple, and low-cost assay. This approach showed great potential in the application of foodborne pathogen detection concerning food safety.


Assuntos
Compostos Férricos/química , Microbiologia de Alimentos/métodos , Listeria monocytogenes/isolamento & purificação , Anticorpos Antibacterianos/química , Anticorpos Imobilizados/química , Coloides , Compostos Férricos/síntese química , Imunoensaio , Limite de Detecção , Listeria monocytogenes/imunologia , Nanoestruturas/química , Fitas Reagentes/química
19.
Epidemiol Infect ; 149: e220, 2021 09 30.
Artigo em Inglês | MEDLINE | ID: mdl-34588077

RESUMO

A case of listeriosis occurred in a hospitalised patient in England in July 2017. Analysis by whole genome sequencing of the Listeria monocytogenes from the patient's blood culture was identified as clonal complex (CC) 121. This culture was indistinguishable to isolates from sandwiches, salads and the maufacturing environment of Company X which supplied these products widely to the National Health Service. Whilst an inpatient, the case was served sandwiches produced by this company on 12 occasions. No other cases infected by this type were detected in the UK between 2016 and 2020. Between 2016 and 2020, more than 3000 samples of food, food ingredients and environmental swabs from this company were tested. Listeria monocytogenes contamination rates declined after July 2017 from 31% to 0.3% for salads and 3% to 0% for sandwiches. A monophyletic group of 127 L. monocytogenes CC121 isolates was recovered during 2016-2019 and was used to estimate the time of the most recent common ancestor as 2014 (95% CI of between 2012 and 2016). These results represent persistent contamination of equipment, food contact surfaces and foods at a food manufacturer by a single L. monocytogenes strain. Colonisation and persistent contamination of food and production environments are risks for public health.


Assuntos
Microbiologia de Alimentos/estatística & dados numéricos , Serviço Hospitalar de Nutrição , Listeria monocytogenes/isolamento & purificação , Listeriose/etiologia , Inglaterra , Manipulação de Alimentos/normas , Doenças Transmitidas por Alimentos/epidemiologia , Doenças Transmitidas por Alimentos/etiologia , Humanos , Listeria monocytogenes/genética , Listeriose/epidemiologia , Masculino , Pessoa de Meia-Idade , Sequenciamento Completo do Genoma
20.
BMC Infect Dis ; 21(1): 564, 2021 Jun 12.
Artigo em Inglês | MEDLINE | ID: mdl-34118865

RESUMO

BACKGROUND: Listeria monocytogenes (LM) has come to be a major public health issue of at-risk groups, causing high morbidity and mortality. Despite this data, studies are very limited in developing countries like Ethiopia. Thus, we aimed to isolate and characterize LM in terms of antibiogram and biofilm formation among pregnant women with fever, women with a history of spontaneous abortion, women with a history of fetal loss, and women with preterm delivery at Jimma University Medical Center (JUMC), southwest Ethiopia. METHODS: A cross-sectional study was done among 144 women from June to August 2019. Isolates were tested for antibiotic susceptibility and biofilm formation using disc diffusion and microtiter plate method, respectively. Data were collected using a structured questionnaire, entered into Epidata 3.1 and logistic regression was done by SPSS v25.0. RESULTS: LM was isolated in 8 (5.56%) of 144 screened women. The isolation rate of LM was relatively higher among women with a history of fetal loss (9.7%), followed by women with preterm delivery (6.25%). One of the six cord blood was positive for LM, indicating that the transplacental transmission rate at JUMC was 16.7%. More than 2% of women with an ongoing pregnancy were found to have LM septicemia, which could hurt their fetus. All of the isolates tested were susceptible to Ampicillin. However, all of the isolates were resistant to Penicillin and Meropenem and were biofilm producers. CONCLUSIONS: The high magnitude of pregnancy-related listeriosis in the current study setting appears that implementation of educational programs targeting risk reduction and more studies to identify sources of LM are warranted. The choice of antibiotics should be after susceptibility testing.


Assuntos
Antibacterianos/farmacologia , Listeria monocytogenes , Listeriose/microbiologia , Complicações Infecciosas na Gravidez/microbiologia , Centros Médicos Acadêmicos , Estudos Transversais , Etiópia/epidemiologia , Feminino , Humanos , Listeria monocytogenes/efeitos dos fármacos , Listeria monocytogenes/isolamento & purificação , Listeriose/prevenção & controle , Testes de Sensibilidade Microbiana , Gravidez , Complicações Infecciosas na Gravidez/prevenção & controle
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