Rapid real-time loop-mediated isothermal amplification combined with coated activated carbon for detection of low numbers of Salmonella enterica from lettuce without enrichment

Wu, Guo Ping; Chen, Su Hua; Levin, Robert E.

doi:10.1016/j.foodcont.2015.03.008

Cited by 16 publications

(7 citation statements)

References 17 publications

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“…Moreover, a standard curve with a high correlation coefficient was obtained in this study, as shown in Fig. 3B , indicating that except for microorganism quantification 32 33 34 , the RT-LAMP assay can also be applied in human DNA quantification. Therefore, the RT-LAMP method can be used for the determination of trace amounts of DNA of interest among copious background DNA, such as specific mutation detection in circulating tumour DNA 24 35 , and can be applied for complex gene quantification, which is clinically meaningful 36 37 .…”

Section: Discussionsupporting

confidence: 58%

Establishment and application of a real-time loop-mediated isothermal amplification system for the detection of CYP2C19 polymorphisms

Zhang

Yao²,

Zhu³

et al. 2016

Sci Rep

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Single-nucleotide polymorphisms (SNPs) represent the most widespread type of genetic variation (approximately 90%) in the human genome, and the demand to overcome such variation has received more attention now than ever before. The capacity to rapidly assess SNPs that correlate with disease predisposition, drug efficacy and drug toxicity is a key step for the development of personalized medicine. In this work, a rapid one-step SNP detection method, real-time loop-mediated isothermal amplification (RT-LAMP), was first applied for CYP2C19 polymorphisms testing. The optimized method was established with specifically designed primers for target amplification by real-time detection in approximately 30 min under isothermal conditions. RT-LAMP amplified few copies of template to produce significant amounts of product and quantitatively detected human DNA with compatible specificity and sensitivity. The success in the establishment of this RT-LAMP protocol for CYP2C19 polymorphism testing is significant for the extension of this technique for the detection of other SNPs, which will further facilitate the development of personalized medicine.

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Section: Discussionsupporting

confidence: 58%

Establishment and application of a real-time loop-mediated isothermal amplification system for the detection of CYP2C19 polymorphisms

Zhang

Yao²,

Zhu³

et al. 2016

Sci Rep

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“…Different researchers have studied LAMP assays for Salmonella detection in RTE fresh produce like lettuce, tomatoes and milk (Zhang et al ., 2011 ; Yang et al ., 2015 ; Wu et al ., 2015 ). More precisely Zhang et al ( 2011 ) have found the detection limit of L. monocytogenes in lettuce (2 CFU/25g) and tomato (19 CFU/25g).…”

Section: Resultsmentioning

confidence: 99%

Evaluation of two loop-mediated isothermal amplification methods for the detection of Salmonella Enteritidis and Listeria monocytogenes in artificially contaminated ready-to eat fresh products

et al. 2015

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In the present study, the effectiveness of two loop-mediated isothermal amplification (LAMP) assays was evaluated. Samples of romaine lettuce, strawberries, cherry tomatoes, green onions and sour berries were inoculated with known dilutions (100-108 CFU/g of produce) of S. Enteritidis and L. monocytogenes. With LAMP, assay pathogens can be detected in less than 60 min. The limits of detection of S. Enteritidis and L. monocytogenes depended on the food sample tested and on the presence of enrichment step. After enrichment steps, all food samples were found positive even at low initial pathogen levels. The developed LAMP, assays, are expected to become a valuable, robust, innovative, powerful, cheap and fast monitoring tool, which can be extensively used for routine analysis, and screening of contaminated foods by the food industry and the Public Food Health Authorities.

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“…It is well known that activated carbon exhibits a high level of adsorption capacity which can efficiently remove DNA amplification inhibitors while adsorb target bacterial cells, so the surface of active carbon must be blocked enough. Previous studies (Luan & Levin, ; Wu et al, ) have shown that the recovery of foodborne pathogens was significantly improved after bentonite coated activated carbon (BCAC) treatment. Currently, the majority of quality control systems available for the supervision of milk in the food industry use an initial enrichment followed by a detection system such as PCR or LAMP assay (Kokkinos et al, ; Ranjbar et al, ).…”

Section: Introductionmentioning

confidence: 99%

“…Although, there were many rapid detection methods designed based on PCR and LAMP assays for foodborne pathogen, which could be limited in sensitivity by DNA amplification inhibitors and caused false-negative results (Wang, Huo, Ren, & Li, 2010;Wu, Chen, & Levin, 2015). In milk, components such as Ca 2+ , proteinase, fats, and milk proteins may block DNA and shield it from access by polymerase (Soejima et al, 2008;Wilson, 1997).…”

Section: Introductionmentioning

confidence: 99%

“…Previous studies (Luan & Levin, 2008;Wu et al, 2015) have shown that the recovery of foodborne pathogens was significantly improved after bentonite coated activated carbon (BCAC) treatment. Currently, the majority of quality control systems available for the supervision of milk in the food industry use an initial enrichment followed by a detection system such as PCR or LAMP assay (Kokkinos et al, 2014;Ranjbar et al, 2016).…”

Section: Introductionmentioning

confidence: 99%

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Real‐time loop‐mediated isothermal amplification assays combined with ethidium monoazide bromide and bentonite coated activated carbon for rapid and sensitive detection of viable Escherichia coli O157:H7 from milk without enrichment

Zhang

Zhong

Shu

et al. 2019

Journal of Food Safety

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A real‐time loop‐mediated isothermal amplification (Rti‐LAMP) DNA assay was developed for rapid and sensitive detection of viable Escherichia coli O157:H7 in milk by combining with ethidium monoazide bromide (EMA) and bentonite coated activated carbon (BCAC) treatment without enrichment. The assay involved inoculating 25 mL portions of milk with various numbers of E. coli O157:H7. Then each of milk sample was adsorbed with 4 g BCAC to remove DNA amplification inhibitor. The recovered purified bacteria sample was further treated with EMA for discrimination viable from dead cells in Rti‐LAMP assay. The detection limit of viable E. coli O157:H7 was 10 CFU/mL in milk by the BCAC‐EMA‐Rti‐LAMP, and the entire assay could be completed in 5 hr. Twenty‐five raw milk samples were detected by the BCAC‐EMA‐Rti‐LAMP assay, using three methods as controls including E. coli O157:H7 plate culture, BCAC‐Rti‐LAMP, and Rti‐LAMP combined with enrichment at 37°C (E‐Rti‐LAMP). The results showed that the BCAC‐EMA‐Rti‐LAMP had similar accuracy and sensitivity as that of plate culture, as they both detected one sample of viable E. coli O157:H7 positive from 25 fresh raw milk samples. These data strongly suggested that the BCAC‐EMA‐Rti‐LAMP could rapidly and sensitively detect viable E. coli O157:H7 in milk without enrichment. Practical Applications Escherichia coli O157:H7 is one of the foodborne pathogens in milk, which has low dose of infection and high pathogenicity. In milk, components such as Ca2+, proteinase, fats, and milk proteins may block DNA and shield it from access by polymerase. This study was targeting VT2 and Z3276 genes for rapid and sensitive detection of viable E. coli O157:H7 in milk. BCAC was used to remove DNA amplification inhibitor and improve the detection sensitivity. EMA was used to discriminate viable from dead cells. The assay will be a potential tool in the rapidity, sensitivity, and specificity for detection of E. coli O157:H7 in milk without enrichment.

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Rapid real-time loop-mediated isothermal amplification combined with coated activated carbon for detection of low numbers of Salmonella enterica from lettuce without enrichment

Cited by 16 publications

References 17 publications

Establishment and application of a real-time loop-mediated isothermal amplification system for the detection of CYP2C19 polymorphisms

Establishment and application of a real-time loop-mediated isothermal amplification system for the detection of CYP2C19 polymorphisms

Evaluation of two loop-mediated isothermal amplification methods for the detection of Salmonella Enteritidis and Listeria monocytogenes in artificially contaminated ready-to eat fresh products

Real‐time loop‐mediated isothermal amplification assays combined with ethidium monoazide bromide and bentonite coated activated carbon for rapid and sensitive detection of viable Escherichia coli O157:H7 from milk without enrichment

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