Real-time PCR detection of <i>Listeria monocytogenes</i>
 in infant formula and lettuce following macrophage-based isolation and enrichment

Day, James B.; Basavanna, Uma

doi:10.1111/jam.12674

Cited by 8 publications

(4 citation statements)

References 64 publications

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“…Basically the culture methods require several days to recognize the pathogenic bacteria and to confirm the suspected colonies by biochemical and serological methods (5-7 days). 16,17 The detection of other pathogens can also be time-consuming using conventional methods. In recent decades molecular methods such as real-time PCR (q-PCR) have been used due to their rapid and sensitive detection of pathogenic bacteria in food products and their high accuracy in detecting the presence of a single type of pathogen in food samples.…”

Section: Discussionmentioning

confidence: 99%

Occurrence, Molecular Detection and Antibiotic Resistance Profile of Escherichia coli O157:H7 Isolated from Ready-to-Eat Vegetable Salads in Iran

Kouchakkhani¹,

Dehghan²,

Moosavy³

et al. 2016

Pharm Sci

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Section: Discussionmentioning

confidence: 99%

Occurrence, Molecular Detection and Antibiotic Resistance Profile of Escherichia coli O157:H7 Isolated from Ready-to-Eat Vegetable Salads in Iran

Kouchakkhani¹,

Dehghan²,

Moosavy³

et al. 2016

Pharm Sci

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“…The primer for the detection of L. monocytogenes was designed from the feoB gene sequence. Recent studies concerning virulence-associated genes, such as Imo0460, prs, PrfA, hly, hlyA, iap, flaA, actA, inlA, plcA, plcB, IgY, and inlB, and those involving the pathogenesis of the Listeriosis process have been published [30][31][32]. The previous studies reported the rapid detection of L. monocytogenes foodborne pathogen based on plcB and listeriolysin O (hly) gene using the LAMP-LFD [25] and LAMP-turbidity assays [33], as shown in Table-2 [7,8,16,17,19,20,[21][22][23][24][33][34][35].…”

Section: Discussionmentioning

confidence: 99%

Development of loop-mediated isothermal amplification-lateral flow dipstick as a rapid screening test for detecting Listeria monocytogenes in frozen food products using a specific region on the ferrous iron transport protein B gene

2022

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Background and Aim: Listeria monocytogenes is a critical foodborne pathogen that infects pregnant females and their newborns and older adults and individuals with comorbidities. It contaminates fresh vegetables, fruits, ready-to-eat foods, and frozen food products consumed by individuals. The culture conventional detection methods for L. monocytogenes are time-consuming, taking 4 days. This study aimed to describe the development and comparison of loop-mediated isothermal amplification (LAMP)- lateral flow dipstick (LFD), LAMP assay to PCR, and conventional culture for detecting L. monocytogenes in frozen food products. Materials and Methods: Five LAMP primer sets, including F3, B3, forward inner primer, and backward inner primer, were designed from a specific region on ferrous iron transport protein B gene (feoB gene) to amplify LAMP products. The DNA probe was created, and the detection limit was determined in pure culture and purified DNA, as well as the detection in 20 frozen food product samples. Results: The LMfeoB4 LAMP primer sets and DNA probe were LAMP products amplified at 60°C for 50 min. The specificity of the assay revealed no cross-reactivity with other pathogenic bacteria. The limit of detection (LOD) of the LAMP-LFD and LAMP assays using purified genomic DNA was 219 fg/μL both in LAMP and LAMP-LFD assays. The LOD of LAMP and LAMP-LFD assays in pure culture was 4.3×102 colony-forming unit (CFU)/mL and 43 CFU/mL, respectively. The LOD of the LAMP-LFD assay using artificially inoculated chicken in frozen food samples with pre-enrichment was 3.2×102 CFU/mL. The LAMP-LFD was also more sensitive than the LAMP assay and polymerase chain reaction. Finally, LAMP-LFD revealed no false positives in any of the 20 frozen food product samples. Conclusion: LAMP-LFD assay using a specific region on the feoB gene to detect L. monocytogenes was highly specific, sensitive, faster, and convenient, making it a valuable tool for the monitoring and rapid screening of L. monocytogenes in frozen food products. This technique is applicable to the development of detection technologies for other pathogens in food products.

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“…This technique can be used to detect and quantify bacterial pathogens on contaminated surfaces. It has reported that qPCR has been used to rapidly detect Listeria monocytogenes in raw milk and soft cheese [22], in infant formula and lettuce [23] and pork meat [24]. In order to evaluate the bacterial populations in biofilms, qPCR using fluorescent dyes methods were used to quantify the number of RNA transcripts of specific genes [25].…”

Section: Transcriptomics In Food Microbiologymentioning

confidence: 99%

Foodomics in microbiological investigations

2015

Current Opinion in Food Science

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Real-time PCR detection of Listeria monocytogenes in infant formula and lettuce following macrophage-based isolation and enrichment

Cited by 8 publications

References 64 publications

Occurrence, Molecular Detection and Antibiotic Resistance Profile of Escherichia coli O157:H7 Isolated from Ready-to-Eat Vegetable Salads in Iran

Occurrence, Molecular Detection and Antibiotic Resistance Profile of Escherichia coli O157:H7 Isolated from Ready-to-Eat Vegetable Salads in Iran

Development of loop-mediated isothermal amplification-lateral flow dipstick as a rapid screening test for detecting Listeria monocytogenes in frozen food products using a specific region on the ferrous iron transport protein B gene

Foodomics in microbiological investigations

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