PCR Mediated Nucleic Acid Molecular Recognition Technology for Detection of Viable and Dead Foodborne Pathogens

Chen, Mengtao; Lan, Xinyue; Zhu, Longjiao; Ru, P.; Xu, Wentao; Liu, Haiyan

doi:10.3390/foods11172675

Cited by 20 publications

(8 citation statements)

References 28 publications

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“…Viable PCR (vPCR), another variant of PCR that involves the use of intercalating dyes such as ethidium monoazide (EMA) and propidium monoazide (PMA) for sample pre-treatment followed by PCR amplification allowing for the detection of living foodborne pathogens [ 109 ]. vPCR could be more readily applied for the detection of pathogens than RT PCR.…”

Section: Methods For Detecting Foodborne Pathogensmentioning

confidence: 99%

“…These intercalating dyes can penetrate the membrane of damaged or dead bacterial cells, irreversibly binding to their DNA molecules thereby preventing their amplification by PCR primers. Therefore, any amplicon observed from the subsequent PCR will be from live bacterial cells [ 109 ]. It has been used to detect live Salmonella sp.…”

Section: Methods For Detecting Foodborne Pathogensmentioning

confidence: 99%

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A Review of Modern Methods for the Detection of Foodborne Pathogens

Aladhadh

2023

Microorganisms

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Despite the recent advances in food preservation techniques and food safety, significant disease outbreaks linked to foodborne pathogens such as bacteria, fungi, and viruses still occur worldwide indicating that these pathogens still constitute significant risks to public health. Although extensive reviews of methods for foodborne pathogens detection exist, most are skewed towards bacteria despite the increasing relevance of other pathogens such as viruses. Therefore, this review of foodborne pathogen detection methods is holistic, focusing on pathogenic bacteria, fungi, and viruses. This review has shown that culture-based methods allied with new approaches are beneficial for the detection of foodborne pathogens. The current application of immunoassay methods, especially for bacterial and fungal toxins detection in foods, are reviewed. The use and benefits of nucleic acid-based PCR methods and next-generation sequencing-based methods for bacterial, fungal, and viral pathogens’ detection and their toxins in foods are also reviewed. This review has, therefore, shown that different modern methods exist for the detection of current and emerging foodborne bacterial, fungal, and viral pathogens. It provides further evidence that the full utilization of these tools can lead to early detection and control of foodborne diseases, enhancing public health and reducing the frequency of disease outbreaks.

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Section: Methods For Detecting Foodborne Pathogensmentioning

confidence: 99%

Section: Methods For Detecting Foodborne Pathogensmentioning

confidence: 99%

A Review of Modern Methods for the Detection of Foodborne Pathogens

Aladhadh

2023

Microorganisms

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“…To determine the phylogenetic relationship of A. hydrophila, a phylogenetic tree analysis was conducted on the 16S rRNA and gyrB genes of A. hydrophila. This analysis employed the bootstrap method with 1,000 bootstrap replications, and it was carried out using MEGA 11.0 (Molecular Evolutionary Genetic Analysis), as described by Chen, [64].…”

Section: Amplification Sequencing and Phylogenetic Tree Analysis Of 1...mentioning

confidence: 99%

Molecular characterization of Aeromonas hydrophila detected in Channa marulius and Sperata sarwari sampled from rivers of Punjab in Pakistan

Mahmood,

Rasool,

Hafeez-ur-Rehman

et al. 2024

PLoS ONE

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Aeromonas hydrophila is one of the major pathogenic bacteria responsible for causing severe outbreaks at fish farms and is also a major global public health concern. This bacterium harbors many virulence genes. The current study was designed to evaluate the antidrug and virulence potential of A. hydrophila by amplifying its antimicrobial resistance and virulence genes using PCR and examining their effects on fish tissues and organs. A total of 960 fish samples of Channa marulius and Sperata sarwari were collected from four sites of the rivers of the Punjab, Pakistan. A. hydrophila isolates were subjected to biochemical identification and detection of virulence and antimicrobial resistance (AMR) genes by PCR. We retrieved 181 (6.46%) A. hydrophila isolates from C. marulius and 177 (6.25%) isolates from S. sarwari. Amplification through PCR revealed the incidence of virulence genes in 95.7% of isolates in C. marulius and 94.4% in S. sarwari. Similarly, amplification through PCR also revealed occurrence of AMR genes in 87.1% of isolates in C. marulius and 83.9% in S. sarwari. Histopathological examination revealed congestion (5.2%) and hepatocyte necrosis (4.6%) in liver, lamellar fusion (3.3%) and the presence of bacterial colonies (3.7%) in gills, fin erosion (6%), and the presence of biofilms (3.5%) in tail fins of infected fish. Phylogenetic tree analysis of 16S rRNA and gyrB gene of A. hydrophila revealed 100% and 97% similarity, respectively, with 16S rRNA gene and gyrB of A. hydrophila isolated in previous studies. The results of antimicrobial susceptibility testing showed that all isolates demonstrated resistance to sulfamethoxazole, ampicillin, neomycin, and norfloxacin, while susceptibility to gentamicin, chloramphenicol, and tetracycline, and intermediate resistance was observed against cefotaxime. The results concluded that examined fish samples were markedly contaminated with virulent and multidrug strains of A. hydrophila which may be of a potential health risk. The study emphasizes the responsible antimicrobial use in aquaculture and the urgent need for effective strategies to control the spread of virulence and antimicrobial resistance genes in A. hydrophila.

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“…In addition to efforts on enabling the detection of the viable pathogen in food samples, identification and optimization of other potential viability dyes are also suggested. Previous studies have indicated that DyeTox13 and thiazole orange monoazide (TOMO) potentially allow for the suppression of DNA signals from dead cells ( Chen et al, 2022 , Wang et al, 2022a ). Thus far, however, very little information is available about the effectiveness of these new dyes to inhibit the DNA signals from dead cells present in food samples.…”

Section: Remaining Challenges and Possible Opportunitiesmentioning

confidence: 99%

Rapid detection methods for foodborne pathogens based on nucleic acid amplification: Recent advances, remaining challenges, and possible opportunities

Ndraha,

Lin,

Wang

et al. 2023

Food Chemistry: Molecular Sciences

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PCR Mediated Nucleic Acid Molecular Recognition Technology for Detection of Viable and Dead Foodborne Pathogens

Cited by 20 publications

References 28 publications

A Review of Modern Methods for the Detection of Foodborne Pathogens

A Review of Modern Methods for the Detection of Foodborne Pathogens

Molecular characterization of Aeromonas hydrophila detected in Channa marulius and Sperata sarwari sampled from rivers of Punjab in Pakistan

Rapid detection methods for foodborne pathogens based on nucleic acid amplification: Recent advances, remaining challenges, and possible opportunities

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