Multiplex Recombinase Polymerase Amplification Assay for the Simultaneous Detection of Three Foodborne Pathogens in Seafood

Ma, Bach; Li, Jiali; Chen, Kai; Yu, Xiaoping; Sun, Chuanxin; Zhang, Mingzhou

doi:10.3390/foods9030278

Cited by 50 publications

(46 citation statements)

References 48 publications

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“…Salmonella , one of the main causes of foodborne diseases, is a significant public health concern. Salmonella is known to be spread primarily through the consumption of contaminated food or water [ 1 , 2 , 3 ]. Salmonella infection can cause serious illness in humans, especially infants and the elderly, causing diseases such as typhoid fever and gastroenteritis and even leading to death [ 4 ].…”

Section: Introductionmentioning

confidence: 99%

Rapid Detection of Salmonella Enteritidis, Typhimurium, and Thompson by Specific Peak Analysis Using Matrix-Assisted Laser Desorption Ionization Time-of-Flight Mass Spectrometry

Yang

Kim

et al. 2021

Foods

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Rapid detection of Salmonella serovars is important for the effective control and monitoring of food industries. In this study, we evaluate the application of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry for the rapid detection of three serovars, Enteritidis, Typhimurium, and Thompson, that are epidemiologically important in Korea. All strains were identified at the genus level, with a mean score of 2.319 using the BioTyper database, and their protein patterns were confirmed to be similar by principal component analysis and main spectrum profile dendrograms. Specific peaks for the three serovars were identified by analyzing 65 reference strains representing 56 different serovars. Specific mass peaks at 3018 ± 1 and 6037 ± 1, 7184 ± 1, and 4925 ± 1 m/z were uniquely found in the reference strains of serovars Enteritidis, Typhimurium, and Thompson, respectively, and they showed that the three serovars can be differentiated from each other and 53 other serovars. We verified the reproducibility of these mass peaks in 132 isolates, and serovar classification was achieved with 100% accuracy when compared with conventional serotyping through antisera agglutination. Our method can rapidly detect a large number of strains; hence, it will be useful for the high-throughput screening of Salmonella serovars.

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Section: Introductionmentioning

confidence: 99%

Rapid Detection of Salmonella Enteritidis, Typhimurium, and Thompson by Specific Peak Analysis Using Matrix-Assisted Laser Desorption Ionization Time-of-Flight Mass Spectrometry

Yang

Kim

et al. 2021

Foods

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“…There are usually multiple pathogens in food; thus, traditional single-target nucleic acid detection can no longer meet the needs of multi-target detection. The ability to carry out multiple RPA amplifications in one reaction system has been successfully reported; for example, the RPA assay was used for the simultaneous detection of three food-borne pathogens in seafood and multiple bacteria in urine [ 25 , 34 ]. RPA combined with colloidal gold test strips has also been used, such as for the rapid detection of coliform bacteria using a lateral flow test strip assay [ 35 ], as well as in a dual fluorescein isothiocyanate (FITC) lateral flow immunoassay for the sensitive detection of Escherichia coli O157:H7 in food samples [ 36 ].…”

Section: Discussionmentioning

confidence: 99%

“…The primers (Table 2) with target-specific labels were tagged with cyanine 5 (Cy5) and digoxin (primers for the hlyA gene), biotin and digoxin (primers for the toxR gene), or carboxy fluorescein (FAM) and digoxin (primers for the rfbE gene). The primers of Vibrio parahaemolyticus were obtained from our previous work [25]. RPA was performed in a 50 µL volume with a TwistAmp Basic Kit (TwistDX, Cambridge, UK).…”

Section: Primer Designmentioning

confidence: 99%

A Rapid and Sensitive Europium Nanoparticle-Based Lateral Flow Immunoassay Combined with Recombinase Polymerase Amplification for Simultaneous Detection of Three Food-Borne Pathogens

Chen

et al. 2021

IJERPH

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Food-borne pathogens have become an important public threat to human health. There are many kinds of pathogenic bacteria in food consumed daily. A rapid and sensitive testing method for multiple food-borne pathogens is essential. Europium nanoparticles (EuNPs) are used as fluorescent probes in lateral flow immunoassays (LFIAs) to improve sensitivity. Here, recombinase polymerase amplification (RPA) combined with fluorescent LFIA was established for the simultaneous and quantitative detection of Listeria monocytogenes, Vibrio parahaemolyticus, and Escherichia coliO157:H7. In this work, the entire experimental process could be completed in 20 min at 37 °C. The limits of detection (LODs) of EuNP-based LFIA–RPA were 9.0 colony-forming units (CFU)/mL for Listeria monocytogenes, 7.0 CFU/mL for Vibrio parahaemolyticus, and 4.0 CFU/mL for Escherichia coliO157:H7. No cross-reaction could be observed in 22 bacterial strains. The fluorescent LFIA–RPA assay exhibits high sensitivity and good specificity. Moreover, the average recovery of the three food-borne pathogens spiked in food samples was 90.9–114.2%. The experiments indicate the accuracy and reliability of the multiple fluorescent test strips. Our developed EuNP-based LFIA–RPA assay is a promising analytical tool for the rapid and simultaneous detection of multiple low concentrations of food-borne pathogens.

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“…Like PCR, only a pair of primer is required for RPA reaction, making the assay design relatively easy; however, the multiplexing ability is yet to be fully explored. Notwithstanding, RPA has been used to multiplex food pathogens like Staphylococcus aureus, Vibrio parahaemolyticus and Salmonella enteritidis coupled on a lateral flow strip, and the test line intensities were quantified using a test strip reader [65]. It will be useful to see this recent strategy deployed for SARS-CoV-2 multiplexing as well.…”

Section: Development Of Multiplex Detection Of Sars-cov-2mentioning

confidence: 99%

COVID-19 Infection Diagnosis: Potential Impact of Isothermal Amplification Technology to Reduce Community Transmission of SARS-CoV-2

James

Alawneh

2020

Diagnostics

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The current coronavirus disease 2019 (COVID-19) pandemic is largely driven by community transmission, after 2019 novel Coronavirus (2019-nCoV or SARS-CoV-2) crosses the borders. To stop the spread, rapid testing is required at community clinics and hospitals. These rapid tests should be comparable with the standard PCR technology. Isothermal amplification technology provides an excellent alternative that is highly amenable to resource limited settings, where expertise and infrastructure to support PCR are not available. In this review, we provide a brief description of isothermal amplification technology, its potential and the gaps that need to be considered for SARS-CoV-2 detection. Among this emerging technology, loop-mediated amplification (LAMP), recombinase polymerase amplification (RPA) and Nicking enzyme-assisted reaction (NEAR) technologies have been identified as potential platforms that could be implemented at community level, without samples referral to a centralized laboratory and prolonged turnaround time associated with the standard COVID-19 RT-PCR test. LAMP, for example, has recently been shown to be comparable with PCR and could be performed in less than 30 min by non-laboratory staff, without RNA extractions commonly associated with PCR. Interestingly, NEAR (ID NOW™ COVID-19 (Abbott, IL, USA) was able to detect the virus in 5 min. More so, isothermal platforms are cost effective and could easily be scaled up to resource limited settings. Diagnostics developers, scientific community and commercial companies could consider this alternative method to help stop the spread of COVID-19.

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Multiplex Recombinase Polymerase Amplification Assay for the Simultaneous Detection of Three Foodborne Pathogens in Seafood

Cited by 50 publications

References 48 publications

Rapid Detection of Salmonella Enteritidis, Typhimurium, and Thompson by Specific Peak Analysis Using Matrix-Assisted Laser Desorption Ionization Time-of-Flight Mass Spectrometry

Rapid Detection of Salmonella Enteritidis, Typhimurium, and Thompson by Specific Peak Analysis Using Matrix-Assisted Laser Desorption Ionization Time-of-Flight Mass Spectrometry

A Rapid and Sensitive Europium Nanoparticle-Based Lateral Flow Immunoassay Combined with Recombinase Polymerase Amplification for Simultaneous Detection of Three Food-Borne Pathogens

COVID-19 Infection Diagnosis: Potential Impact of Isothermal Amplification Technology to Reduce Community Transmission of SARS-CoV-2

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