Detection of E. coli O157:H7 in Food Using Automated Immunomagnetic Separation Combined with Real-Time PCR

Park, Ji Young; Lim, Min-Cheol; Park, Kisang; Ok, Gyeongsik; Chang, Hyun-Joo; Lee, Nari; Park, Tae Jung; Choi, Sung‐Wook

doi:10.3390/pr8080908

Cited by 9 publications

(8 citation statements)

References 33 publications

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“… (A) Step-by-step schematic representation of ELISA ( Zuo et al, 2021 ); (B) Schematic illustration of the detection method for E. coli O157:H7 using the automated IMS device combined with real-time PCR ( Park et al, 2020 ). …”

Section: Immunological Detection Techniquesmentioning

confidence: 99%

“…Moreover, Park et al (2020) described the development of an automated IMS device combined with real-time PCR for detecting foodborne bacteria ( Figure 2B ). Target bacteria in the range of 10 1 –10 2 colony-forming units per mg or g of sample can be detected in food samples, such as milk, ground beef, and cabbage, by using the proposed approach.…”

Section: Immunological Detection Techniquesmentioning

confidence: 99%

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Research progress on detection techniques for point-of-care testing of foodborne pathogens

Zhao

Huang

et al. 2022

Front. Bioeng. Biotechnol.

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The global burden of foodborne disease is enormous and foodborne pathogens are the leading cause of human illnesses. The detection of foodborne pathogenic bacteria has become a research hotspot in recent years. Rapid detection methods based on immunoassay, molecular biology, microfluidic chip, metabolism, biosensor, and mass spectrometry have developed rapidly and become the main methods for the detection of foodborne pathogens. This study reviewed a variety of rapid detection methods in recent years. The research advances are introduced based on the above technical methods for the rapid detection of foodborne pathogenic bacteria. The study also discusses the limitations of existing methods and their advantages and future development direction, to form an overall understanding of the detection methods, and for point-of-care testing (POCT) applications to accurately and rapidly diagnose and control diseases.

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Section: Immunological Detection Techniquesmentioning

confidence: 99%

Section: Immunological Detection Techniquesmentioning

confidence: 99%

Research progress on detection techniques for point-of-care testing of foodborne pathogens

Zhao

Huang

et al. 2022

Front. Bioeng. Biotechnol.

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“…Furthermore, MFGs and proteins such as caseins can disrupt the interaction between immunomagnetic beads and STEC during the confirmation step. Tween 20 can be added at this stage to improve sample homogenization and block non-specific interactions [ 224 ].…”

Section: Consequences Of the Stec–mfg Associationmentioning

confidence: 99%

Shiga Toxin-Producing Escherichia coli and Milk Fat Globules

Bagel

Sergentet

2022

Microorganisms

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Shiga toxin-producing Escherichia coli (STEC) are zoonotic Gram-negative bacteria. While raw milk cheese consumption is healthful, contamination with pathogens such as STEC can occur due to poor hygiene practices at the farm level. STEC infections cause mild to serious symptoms in humans. The raw milk cheese-making process concentrates certain milk macromolecules such as proteins and milk fat globules (MFGs), allowing the intrinsic beneficial and pathogenic microflora to continue to thrive. MFGs are surrounded by a biological membrane, the milk fat globule membrane (MFGM), which has a globally positive health effect, including inhibition of pathogen adhesion. In this review, we provide an update on the adhesion between STEC and raw MFGs and highlight the consequences of this interaction in terms of food safety, pathogen detection, and therapeutic development.

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“…The limit of detection (LOD) for the colony-forming units in most foods in the food hygiene inspection standards is not very high. Moreover, the pre-processing of many detection methods are time-consuming and laborious, requiring multiple subcultures and biochemical tests (Park et al, 2020a). Therefore, a method is needed that can rapidly enrich E. coli in samples during the pretreatment stage.…”

Section: Introductionmentioning

confidence: 99%

Outer membrane protein A (OmpA) may be used as a novel target to enrich and detect Escherichia coli in milk samples

Tian

Yang²,

Shi

et al. 2022

Journal of Dairy Science

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In recent years, food safety incidents caused by Escherichia coli have occurred frequently, bringing a serious endangerment to human health. Due to the complex matrix of milk samples and the long pretreatment time, the existing methods cannot quickly detect E. coli in milk samples. It is necessary to enrich the E. coli in the complex matrix to improve the detection sensitivity. The E. coli outer membrane protein A (OmpA) is widely present on the cell membrane of E. coli and may be used as a new target to enrich E. coli. In this study, the purified recombinant OmpA protein was used to immunize BALB/c mice to produce polyclonal antibody. Immunomagnetic beads were combined with the polyclonal antibody to enrich the E. coli in the artificially contaminated milk samples. The products of immunoprecipitation were further used for PCR assay. The bacteria in the PCR sample can be pre-enriched, and the limit of detection is 10 × 10 0 cfu/mL, which is about 100 times more sensitive than samples not processed by this method. Then, the artificially contaminated milk, coffee, juice, and soybean milk samples were tested separately, and it was found that the E. coli gene could be amplified. The whole analysis time was about 120 min, including the enrichment of bacteria and the detection of eluate. We found that OmpA combined with immunomagnetic beads was more efficient, fast, and convenient than the conventional method. Bacteria can be enriched more efficiently without extracting genomic DNA and culturing bacteria. Therefore, this method has potential value for improving the detection sensitivity and shortening the detection time of E. coli in food samples.

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Detection of E. coli O157:H7 in Food Using Automated Immunomagnetic Separation Combined with Real-Time PCR

Cited by 9 publications

References 33 publications

Research progress on detection techniques for point-of-care testing of foodborne pathogens

Research progress on detection techniques for point-of-care testing of foodborne pathogens

Shiga Toxin-Producing Escherichia coli and Milk Fat Globules

Outer membrane protein A (OmpA) may be used as a novel target to enrich and detect Escherichia coli in milk samples

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