Development of a Rapid Screening Test for Listeria monocytogenes in Raw Chicken Meat Using Loop-Mediated Isothermal Amplification (LAMP) and Lateral Flow Dipstick (LFD)

Wachiralurpan, Sirirat; Sriyapai, Thayat; Areekit, Supatra; Kaewphinit, Thongchai; Sriyapai, Pichapak; Santiwatanakul, Somchai; Chansiri, Kosum

doi:10.1007/s12161-017-0949-4

Cited by 20 publications

(23 citation statements)

References 34 publications

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“…A single colony of bacterial strain on brain heart infusion (BHI) agar (BBL, Becton Dickinson Microbiology Systems, Cockeysville, MD, USA) was cultured in BHI broth with constant shaking (250 rpm) at 37°C overnight. The extracted DNAs of all strains were obtained as previously described method (Wachiralurpan et al, 2017a ). The rapid DNA extraction was performed by suspension of bacterial cell pellets in 100 μL of sterile deionized, nanopure water prior to boiling at 100°C for 10 min.…”

Section: Methodsmentioning

confidence: 99%

“…According to the previous report (Wachiralurpan et al, 2017a ), a plcB primers set and DNA probe (Petty patent submission number 1601004756) were commercially synthesized by Bio Basic Inc., Canada. The LAMP and PCR reactions were performed according to the methods described in previous report (Wachiralurpan et al, 2017a ). The negative controls were included in each DNA amplification run.…”

Section: Methodsmentioning

confidence: 99%

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Rapid Colorimetric Assay for Detection of Listeria monocytogenes in Food Samples Using LAMP Formation of DNA Concatemers and Gold Nanoparticle-DNA Probe Complex

et al. 2018

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Listeria monocytogenes is a major foodborne pathogen of global health concern. Herein, the rapid diagnosis of L. monocytogenes has been achieved using loop-mediated isothermal amplification (LAMP) based on the phosphatidylcholine-phospholipase C gene (plcB). Colorimetric detection was then performed through the formation of DNA concatemers and a gold nanoparticle/DNA probe complex (GNP/DNA probe). The overall detection process was accomplished within approximately 1 h with no need for complicated equipment. The limits of detection for L. monocytogenes in the forms of purified genomic DNA and pure culture were 800 fg and 2.82 CFU mL−1, respectively. No cross reactions were observed from closely related bacteria species. The LAMP-GNP/DNA probe assay was applied to the detection of 200 raw chicken meat samples and compared to routine standard methods. The data revealed that the specificity, sensitivity, and accuracy were 100, 90.20, and 97.50%, respectively. The present assay was 100% in conformity with LAMP-agarose gel electrophoresis assay. Five samples that were negative by both assays appeared to have the pathogen at below the level of detection. The assay can be applied as a rapid direct screening method for L. monocytogenes.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Rapid Colorimetric Assay for Detection of Listeria monocytogenes in Food Samples Using LAMP Formation of DNA Concatemers and Gold Nanoparticle-DNA Probe Complex

et al. 2018

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“…Referring to our previous publication, the optimum LAMP was achieved at 60 min. 21 As the LAMP reaction progresses, pyrophosphate ions produced from deoxyribonucleotides (dNTPs) bind to magnesium ions and subsequently form a white precipitate of magnesium pyrophosphate. 22 The amount of the precipitate can be measured using a real-time turbidimeter, which is more appropriate for monitoring LAMP reactions because it can detect turbidity at low levels and presents no issues with aerosol contamination.…”

Section: Discussionmentioning

confidence: 99%

“…According to our previous publication, the hly-PCR assay was 10 times more sensitive than the hly-LAMP assay. 21 It is possible that the 32 samples of both LAMP-turbidity and PCR positive contain bacteria in the range of detection whereas 4 real samples of discrepancy between the two techniques (PCR positive, LAMP negative) may contain the amount of DNA less than the limit of detection of hly-LAMP. However, it was suggested that the pre-enrichment sample prior to the LAMP-turbidity assay can improve the sensitivity of the test.…”

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confidence: 99%

A one-step rapid screening test ofListeria monocytogenesin food samples using a real-time loop-mediated isothermal amplification turbidity assay

et al. 2017

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A rapid and specific, hly-based, loop-mediated isothermal amplification (LAMP) was applied for the detection of Listeria monocytogenes in food and food products, using a real-time turbidimeter platform (LAMP-turbidity). The principle behind this method relies on an increase in a DNA yield, which correlates with the production of magnesium pyrophosphate, and the results can be determined via an amplification curve within 1 h. The specificity test revealed that L. monocytogenes (DMST 17303) was observed from 34.1 to 38.3 min, while thirty strains of non-L. monocytogenes demonstrated no crossreactions. The limits of detection for purified genomic DNA and pure culture were 800 pg mL À1 and 2.82Â 10 3 CFU mL À1 , respectively. Investigation on 200 raw chicken meat samples indicated that the specificity, sensitivity, and accuracy of LAMP-turbidity were 100%, 62.75%, and 90.50%, respectively.These data suggest that an hly-based, real-time, quantitative LAMP-turbidity assay can be an applicable tool for the epidemiological screening of L. monocytogenes in food and food products.

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“…The LAMP technique is a novel constant-temperature nucleic acid amplification technique invented by Japanese scholars in 2000 with high sensitivity, specificity, and rapidity for the low-cost detection of pathogens (13)(14)(15)(16)(17). Song found that a loop-mediated isothermal amplification (LAMP) method for detecting Shigella and enteroinvasive Escherichia coli, the LAMP method efficiently detected the gene within 2 h at a minimal amount of bacteria (8 CFU) per reaction (18).…”

Section: Introductionmentioning

confidence: 99%

A multiplex loop-mediated isothermal amplification assay for rapid detection of <i>Bacillus cereus</i> and <i>Staphylococcus aureus </i>

Deng

Liu²,

Jiang

et al. 2019

BST

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Bacillus cereus (B. cereus) and Staphylococcus aureus (S. aureus) are major human foodborne pathogens that may produce a variety of toxins and cause diarrhea, food poisoning, and even death. In order to monitor and prevent the spread of these pathogens, a multiplex loop-mediated isothermal amplification (multi-LAMP) assay was developed to simultaneously and rapidly detect B. cereus and S. aureus. The sensitivity and specificity of the loop-mediated isothermal amplification (LAMP) reactions were determined via electrophoresis. The multi-LAMP showed 100% inclusivity and exclusivity, the sensitivity was 10 fg/μL and was 10 times more sensitive than that of polymerase chain reaction (PCR), the results were consistent with those of conventional PCR assay, and the entire assay should be finished within 40 min. This multi-LAMP assay was confirmed as a rapid and reliable diagnostic technique upon application for clinical samples and food samples. To our knowledge, this is the first study to report the application of multi-LAMP to detect B. cereus and S. aureus.

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Development of a Rapid Screening Test for Listeria monocytogenes in Raw Chicken Meat Using Loop-Mediated Isothermal Amplification (LAMP) and Lateral Flow Dipstick (LFD)

Cited by 20 publications

References 34 publications

Rapid Colorimetric Assay for Detection of Listeria monocytogenes in Food Samples Using LAMP Formation of DNA Concatemers and Gold Nanoparticle-DNA Probe Complex

Rapid Colorimetric Assay for Detection of Listeria monocytogenes in Food Samples Using LAMP Formation of DNA Concatemers and Gold Nanoparticle-DNA Probe Complex

A one-step rapid screening test ofListeria monocytogenesin food samples using a real-time loop-mediated isothermal amplification turbidity assay

A multiplex loop-mediated isothermal amplification assay for rapid detection of <i>Bacillus cereus</i> and <i>Staphylococcus aureus </i>

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