scholarly journals A Rapid and Sensitive Europium Nanoparticle-Based Lateral Flow Immunoassay Combined with Recombinase Polymerase Amplification for Simultaneous Detection of Three Food-Borne Pathogens

2021 ◽  
Vol 18 (9) ◽  
pp. 4574
Author(s):  
Kai Chen ◽  
Biao Ma ◽  
Jiali Li ◽  
Erjing Chen ◽  
Ying Xu ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Vibrio Parahaemolyticus ◽  
Pathogenic Bacteria ◽  
Simultaneous Detection ◽  
Lateral Flow ◽  
Recombinase Polymerase Amplification ◽  
Quantitative Detection ◽  
Bacterial Strains ◽  
Food Borne Pathogens ◽  
Food Borne

Food-borne pathogens have become an important public threat to human health. There are many kinds of pathogenic bacteria in food consumed daily. A rapid and sensitive testing method for multiple food-borne pathogens is essential. Europium nanoparticles (EuNPs) are used as fluorescent probes in lateral flow immunoassays (LFIAs) to improve sensitivity. Here, recombinase polymerase amplification (RPA) combined with fluorescent LFIA was established for the simultaneous and quantitative detection of Listeria monocytogenes, Vibrio parahaemolyticus, and Escherichia coliO157:H7. In this work, the entire experimental process could be completed in 20 min at 37 °C. The limits of detection (LODs) of EuNP-based LFIA–RPA were 9.0 colony-forming units (CFU)/mL for Listeria monocytogenes, 7.0 CFU/mL for Vibrio parahaemolyticus, and 4.0 CFU/mL for Escherichia coliO157:H7. No cross-reaction could be observed in 22 bacterial strains. The fluorescent LFIA–RPA assay exhibits high sensitivity and good specificity. Moreover, the average recovery of the three food-borne pathogens spiked in food samples was 90.9–114.2%. The experiments indicate the accuracy and reliability of the multiple fluorescent test strips. Our developed EuNP-based LFIA–RPA assay is a promising analytical tool for the rapid and simultaneous detection of multiple low concentrations of food-borne pathogens.

Application of DNA microarray technology for detection, identification, and characterization of food-borne pathogens

2006 ◽  
Vol 52 (1) ◽  
pp. 1-8 ◽  
Author(s):  
M Kostrzynska ◽  
A Bachand
Keyword(s):  
Dna Microarray ◽  
Dna Sequences ◽  
Pathogenic Bacteria ◽  
Dna Microarrays ◽  
Simultaneous Detection ◽  
Microarray Technology ◽  
Food Borne Pathogens ◽  
Food Borne ◽  
Target Dna

DNA microarrays represent the latest advance in molecular technology. In combination with bioinformatics, they provide unparalleled opportunities for simultaneous detection of thousands of genes or target DNA sequences and offer tremendous potential for studying food-borne microorganisms. This review provides an up-to-date look at the application of DNA microarray technology to detect food-borne pathogenic bacteria, viruses, and parasites. In addition, it covers the advantages of using microarray technology to further characterize microorganisms by providing information for specific identification of isolates, to understand the pathogenesis based on the presence of virulence genes, and to indicate how new pathogenic strains evolved epidemiologically and phylogenetically.Key words: DNA microarrays, food-borne pathogens, detection.

scholarly journals Multiplex Recombinase Polymerase Amplification Assay for the Simultaneous Detection of Three Foodborne Pathogens in Seafood

Foods ◽  
2020 ◽  
Vol 9 (3) ◽  
pp. 278 ◽  
Author(s):  
Biao Ma ◽  
Jiali Li ◽  
Kai Chen ◽  
Xiaoping Yu ◽  
Chuanxin Sun ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Foodborne Pathogens ◽  
Vibrio Parahaemolyticus ◽  
Salmonella Enteritidis ◽  
Simultaneous Detection ◽  
Test Strip ◽  
Food Samples ◽  
Recombinase Polymerase Amplification ◽  
Quantitative Detection ◽  
Test Strip Reader

Foodborne pathogens can cause foodborne illness. In reality, one food sample may carry more than one pathogen. A rapid, sensitive, and multiple target method for bacteria detection is crucial in food safety. For the simultaneous detection of Staphylococcus aureus, Vibrio parahaemolyticus, and Salmonella Enteritidis, multi-objective recombinase polymerase amplification (RPA) combined with a lateral flow dipstick (LFD) was developed in this study. The whole process, including amplification and reading, can be completed in 15 min at 37 °C. The detection limits were 2.6 × 101 CFU/mL for Staphylococcus aureus, 7.6 × 101 CFU/mL for Vibrio parahaemolyticus, and 1.29 × 101 CFU/mL for Salmonella Enteritidis. Moreover, colored signal intensities on test lines were measured by a test strip reader to achieve quantitative detection for Staphylococcus aureus (R2 = 0.9903), Vibrio parahaemolyticus (R2 = 0.9928), and Salmonella Enteritidis (R2 = 0.9945). In addition, the method demonstrated good recoveries (92.00%–107.95%) in the testing of spiked food samples. Therefore, the multiplex LFD-RPA assay is a feasible method for the rapid, sensitive, and quantitative detection of bacterial pathogens in seafood.

Establishment of Multiple-PCR for Diagnosing Three Food-Borne Pathogenic Bacteria in Aquatic Products

2012 ◽  
Vol 554-556 ◽  
pp. 1029-1032
Author(s):  
Hui Li Xia ◽  
Hui Zheng ◽  
Jin Jun Zhang
Keyword(s):  
Listeria Monocytogenes ◽  
Multiplex Pcr ◽  
Vibrio Parahaemolyticus ◽  
Pathogenic Bacteria ◽  
Pcr Amplification ◽  
National Standard ◽  
Aquatic Products ◽  
Gene Coding ◽  
Food Borne ◽  
Pcr Method

A rapid multiplex-PCR method was established in order to detect three foodborne pathogenic bacteria including Salmonella typli，Vibrio parahaemolyticus and Listeria monocytogenes in aquatic products. Three pairs of oligonucleotide primers were designed for multiplex-PCR amplification according to gene coding invasion protein A of Salmonella typli，gene coding immune response of Vibrio parahaemolyticus and regulation histone gene of Listeria monocytogenes. The amplified fragment sizes of these three bacteria were 213 bp, 369 bp and 517bp, respectively．The specificity of the multiplex- PCR was high and the minimum detection limit reached 103 CFU/mL. The multiplex-PCR method was used to analyze three aquatic product samples compared with national standard methods, the coincidence rate of two methods reached 100%. The method developed in this study had high sensitivity and specificity, which could be applied for the rapid detection and molecular epidemiology survey of food-borne pathogenic bacteria in aquatic products．

scholarly journals Use of multi-locus sequencing typing as identification method for the food-borne pathogen Listeria monocytogenes: a review

2015 ◽  
Vol 5 (1) ◽  
Author(s):  
Sonia Lamon ◽  
Domenico Meloni ◽  
Simonetta Gianna Consolati ◽  
Anna Mureddu ◽  
Rina Mazzette
Keyword(s):  
Listeria Monocytogenes ◽  
Detection Methods ◽  
Causative Organism ◽  
Intracellular Pathogen ◽  
Bacterial Strains ◽  
New Approach ◽  
Food Borne ◽  
Global Epidemiology ◽  
Selection Of

<em>Listeria monocytogenes</em> is an ubiquitous, intracellular pathogen which has been implicated within the past decade as the causative organism in several outbreaks of foodborne diseases. In this review, a new approach to molecular typing primarily designed for global epidemiology has been described: multi-<em>locus</em> sequencing typing (MLST). This approach is novel, in that it uses data that allow the unambiguous characterization of bacterial strains via the Internet. Our aim is to present the currently available selection of references on <em>L. monocytogenes</em> MLST detection methods and to discuss its use as <em>gold</em> <em>standard</em> to <em>L. monocytogenes</em> subtyping method.

scholarly journals Isolation and selection of bacteria against shrimp pathogenic Vibrio parahaemolyticus from shrimp pond water on Duyen Hai district, Tra Vinh province

2020 ◽  
Vol 10 (1) ◽  
pp. 43-52
Author(s):  
Tran Vu Phuong ◽  
Dang Thi Ngoc Thanh ◽  
Cao Ngoc Diep
Keyword(s):  
Vibrio Parahaemolyticus ◽  
Pathogenic Bacteria ◽  
Pond Water ◽  
Shrimp Farming ◽  
Diffusion Method ◽  
Resistant Bacteria ◽  
Bacterial Strains ◽  
Shrimp Pond ◽  
Pathogenic Vibrio ◽  
The Status

Antibiotic has frequently been used in the shrimp-farming process in Vietnam. This leads to the status that antibiotic-resistant bacteria and products do not receive in the market. Bacteria had the resistant ability to pathogenic bacteria in water, and they have an important role in sustainable aquaculture. This study aimed to isolate and select good bacterial strains against Vibrio parahaemolyticus, pathogenic bacteria, on shrimp from 8 samples of shrimp pond water at 3 villages Ngu Lac, Phuoc An and Long Toan of Duyen Hai district, Tra Vinh province on NB agar medium. As a result, fifty-nine bacterial isolates were isolated and 10/59 isolates (16.95%) were identified as resistant to Vibrio parahaemolyticus by the well diffusion method. In 10 isolates, there were 7 isolates had good resistance to select for PCR technique and sequencing. The result indicated that these seven strains, including DH1m, DH2f, DH4d, DH8i, DH8m, DH8n, belonged to Bacilli and DH1n strain belonged to Streptomyces sp.

scholarly journals Searching for Antagonistic Activity of Bacterial Isolates Derived from Food Processing Environments on Some Food-Borne Pathogenic Bacteria

2020 ◽  
Vol 49 (4) ◽  
pp. 415-423
Author(s):  
B. Baráti-Deák ◽  
Cs. Mohácsi-Farkas ◽  
Á. Belák
Keyword(s):  
Escherichia Coli ◽  
Food Processing ◽  
Pathogenic Bacteria ◽  
Inhibitory Effect ◽  
Antagonistic Activity ◽  
Bacterial Isolates ◽  
Bacterial Strains ◽  
Food Borne ◽  
Time Period ◽  
Growth Decline

Bacterial strains with inhibitory effect on Salmonella Hartford, Listeria monocytogenes, Yersinia enterocolitica, and Escherichia coli, respectively, were isolated. Out of the 64 bacteria originated from food processing environments, 20 could inhibit at least one of the tested pathogens, and it was proved that growth decline of the pathogenic bacteria was more remarkable by co-culturing than by using cell-free supernatants of the isolates. Seven different genera (Pseudomonas, Bacillus, Paenibacillus, Macrococcus, Staphylococcus, Serratia, and Rothia) reduced the pathogens’ growth during the time period of analysis, and the strongest inhibitory effect was observed after 24 h between 15 and 30 °C. Sensitivity of the tested human pathogenic bacteria against the inhibitory strains was distinct, as Y. enterocolitica could be inhibited by numerous isolates, while S. Hartford proved to be the most resistant. Our results reveal that the isolated bacteria or their excreted metabolites could hinder pathogen growth when used in sufficient quantities.

scholarly journals Viable-but-NonculturableListeria monocytogenesandSalmonella entericaSerovar Thompson Induced by Chlorine Stress Remain Infectious

mBio ◽  
2018 ◽  
Vol 9 (2) ◽  
pp. e00540-18 ◽  
Author(s):  
Callum J. Highmore ◽  
Jennifer C. Warner ◽  
Steve D. Rothwell ◽  
Sandra A. Wilks ◽  
C. William Keevil
Keyword(s):  
Listeria Monocytogenes ◽  
Caenorhabditis Elegans ◽  
Life Span ◽  
Fresh Produce ◽  
Environmental Stresses ◽  
Vbnc State ◽  
Viable But Nonculturable ◽  
Content Type ◽  
Food Borne Pathogens ◽  
Food Borne

ABSTRACTThe microbiological safety of fresh produce is monitored almost exclusively by culture-based detection methods. However, bacterial food-borne pathogens are known to enter a viable-but-nonculturable (VBNC) state in response to environmental stresses such as chlorine, which is commonly used for fresh produce decontamination. Here, complete VBNC induction of green fluorescent protein-taggedListeria monocytogenesandSalmonella entericaserovar Thompson was achieved by exposure to 12 and 3 ppm chlorine, respectively. The pathogens were subjected to chlorine washing following incubation on spinach leaves. Culture data revealed that total viableL. monocytogenesandSalmonellaThompson populations became VBNC by 50 and 100 ppm chlorine, respectively, while enumeration by direct viable counting found that chlorine caused a <1-log reduction in viability. The pathogenicity of chlorine-induced VBNCL. monocytogenesandSalmonellaThompson was assessed by usingCaenorhabditis elegans. Ingestion of VBNC pathogens byC. elegansresulted in a significant life span reduction (P= 0.0064 andP< 0.0001), and no significant difference between the life span reductions caused by the VBNC and culturableL. monocytogenestreatments was observed.L. monocytogeneswas visualized beyond the nematode intestinal lumen, indicating resuscitation and cell invasion. These data emphasize the risk that VBNC food-borne pathogens could pose to public health should they continue to go undetected.IMPORTANCEMany bacteria are known to enter a viable-but-nonculturable (VBNC) state in response to environmental stresses. VBNC cells cannot be detected by standard laboratory culture techniques, presenting a problem for the food industry, which uses these techniques to detect pathogen contaminants. This study found that chlorine, a sanitizer commonly used for fresh produce, induces a VBNC state in the food-borne pathogensListeria monocytogenesandSalmonella enterica. It was also found that chlorine is ineffective at killing total populations of the pathogens. A life span reduction was observed inCaenorhabditis elegansthat ingested these VBNC pathogens, with VBNCL. monocytogenesas infectious as its culturable counterpart. These data show that VBNC food-borne pathogens can both be generated and avoid detection by industrial practices while potentially retaining the ability to cause disease.

scholarly journals Quantitative Detection of Listeria monocytogenes and Listeria innocua by Real-Time PCR: Assessment of hly, iap, and lin02483 Targets and AmpliFluor Technology

2004 ◽  
Vol 70 (3) ◽  
pp. 1366-1377 ◽  
Author(s):  
David Rodr�guez-L�zaro ◽  
Marta Hern�ndez ◽  
Mariela Scortti ◽  
Teresa Esteve ◽  
Jos� A. V�zquez-Boland ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Real Time ◽  
Real Time Pcr ◽  
Target Genes ◽  
Dynamic Range ◽  
Significant Proportion ◽  
Quantitative Detection ◽  
Target Sequence ◽  
Food Borne ◽  
Highly Sensitive

ABSTRACT We developed and assessed real-time PCR (RTi-PCR) assays for the detection and quantification of the food-borne pathogen Listeria monocytogenes and the closely related nonpathogenic species L. innocua. The target genes were hly and iap for L. monocytogenes and lin02483 for L. innocua. The assays were 100% specific, as determined with 100 Listeria strains and 45 non-Listeria strains, and highly sensitive, with detection limits of one target molecule in 11 to 56% of the reactions with purified DNA and 3 CFU in 56 to 89% of the reactions with bacterial suspensions. Quantification was possible over a 5-log dynamic range, with a limit of 15 target molecules and R 2 values of >0.996. There was an excellent correspondence between the predicted and the actual numbers of CFU in the samples (deviations of <23%). The hly-based assay accurately quantified L. monocytogenes in all of the samples tested. The iap-based assay, in contrast, was unsuitable for quantification purposes, underestimating the bacterial counts by 3 to 4 log units in a significant proportion of the samples due to serovar-related target sequence variability. The combination of the two assays enabled us to classify L. monocytogenes isolates into one of the two major phylogenetic divisions of the species, I and II. We also assessed the new AmpliFluor technology for the quantitative detection of L. monocytogenes by RTi-PCR. The performance of this system was similar to that of the TaqMan system, although the former system was slightly less sensitive (detection limit of 15 molecules in 45% of the reactions) and had a higher quantification limit (60 molecules).

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