Detection of chicken adulteration in beef via ladder-shape melting temperature isothermal amplification (LMTIA) assay

Wang, Yongzhen; Wang, Borui; Wang, Deguo

doi:10.1080/13102818.2022.2081514

Cited by 6 publications

(2 citation statements)

References 33 publications

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“…Waste oil contains many carcinogenic and disease-causing substances and is one of the common quality and safety problems in the production and processing of edible oils. LMTIA assay for meat germplasm has been established in the laboratory, 21,22 and the established LMTIA technique was used to determine whether soybean oil gDNA extracted in Section 3.5.1 contains adulterated components (beef, chicken, mutton, and pork) and the examination result is shown in Fig. 6.…”

Section: Results and Analysismentioning

confidence: 99%

Study on detection of soybean components in edible oil with ladder-shape melting temperature isothermal amplification (LMTIA) assay

Xiao

Wang

et al. 2023

Anal. Methods

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Section: Results and Analysismentioning

confidence: 99%

Study on detection of soybean components in edible oil with ladder-shape melting temperature isothermal amplification (LMTIA) assay

Xiao

Wang

et al. 2023

Anal. Methods

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“…Additionally, the technique is easy to operate and presented with good consistency. LMTIA is currently applied for food adulteration [ 19 , 20 , 21 ] detection and virus detection [ 22 ], while its application in foodborne pathogen detection is still largely unexplored.…”

Section: Introductionmentioning

confidence: 99%

Detection of Listeria monocytogenes in Food Using the Proofman-LMTIA Assay

Song

Wang

et al. 2023

Molecules

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Microbial factors, including bacteria, viruses, and other pathogens, are significant contributors to foodborne illnesses, posing serious food safety risks due to their potential for rapid growth and contamination. Listeria monocytogenes is one of the most common types of foodborne bacteria that can cause serious foodborne diseases or even fatalities. In this study, a novel nucleic acid amplification method called Proofman-LMTIA was employed to detect Listeria monocytogenes contamination in food. This method combines proofreading enzyme-mediated probe cleavage with ladder-shape melting temperature isothermal amplification. A positive recombinant plasmid was used as a control to ensure the accuracy of the detection results, and primers and Proofman probes were specifically designed for the LMTIA. Genomic DNA was extracted, the reaction temperature was optimized, and the primers’ specificity was verified using foodborne pathogens like Staphylococcus aureus, Escherichia coli O157:H7, and Salmonella. The sensitivity was assessed by testing serial dilutions of genomic DNA, and the method’s applicability was confirmed by detecting artificially contaminated fresh pork. The established LMTIA method exhibited both high specificity and sensitivity. At the optimal reaction temperature of 63 °C, the primers specifically identified Listeria monocytogenes contamination in pork at a concentration of 8.0 ± 0.7 colony-forming units (CFUs) per 25 g. Furthermore, the Proofman-LMTIA method was applied to test Listeria monocytogenes DNA in 30 food samples purchased from a Chinese retail market, and reassuringly, all results indicated no contamination. Proofman-LMTIA can serve as a reliable and rapid method for detecting Listeria monocytogenes in food, contributing to public health by safeguarding consumers from foodborne illnesses, and strengthening food safety regulations.

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Comparison of Plant Genomic DNA Extraction Kits Using the Proofman‐LMTIA Amplification Assay

Yao,

Wang,

Zhou

et al. 2024

Phytochemical Analysis

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IntroductionThe extraction of DNA is the basis of molecular biology research. The quality of the extracted DNA is one of the key factors for the success of molecular biology experiments.ObjectiveTo select a suitable DNA extraction method for Chinese medicinal herbs and seeds.MethodsIn this experiment, four commercial DNA extraction kits were used to extract the genomic DNA (gDNA) from the Pinellia ternata (Thunb.) Breit. powder; Arisaema amurense Maxim. powder as well as the seeds of Glycine max (L.) Merr. On the one hand, the concentration and purity of DNA extracted by these four kits were compared. On the other hand, nucleic acid amplification experiments were performed on three samples extracted by each of the four kits by Proofman‐LMTIA methods, which is a novel nucleic acid isothermal amplification technique. The concentration and purity of DNA extracted by different kits were used to determine which methods were suitable for the dry powder of Chinese herbal medicines and seeds. The efficiency of the amplification curve to show whether the extracted DNA can be used in nucleic acid amplification experiments.ResultsThe results showed that the Proofman‐LMTIA methods were of high specificity and the optimal reaction temperatures were 63, 59, and 59°C for P. ternata (Thunb.) Makino; A. amurense Maxim. and G. max (L.) Merr., respectively. The concentration and purity of the gDNAs extracted with all kits were within the acceptable ranges; meanwhile, the amplification of the gDNA extracted by Kit II was of the highest efficiency.ConclusionIn this experiment, the principle, concentration, purity, and time taken for extracting DNA with four kits were compared. The automated extraction kit based on the magnetic method is suitable for extracting DNA from Chinese medicinal herbs and seeds. The extracted DNA is suitable for nucleic acid amplification detection.

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Detection of chicken adulteration in beef via ladder-shape melting temperature isothermal amplification (LMTIA) assay

Cited by 6 publications

References 33 publications

Study on detection of soybean components in edible oil with ladder-shape melting temperature isothermal amplification (LMTIA) assay

Study on detection of soybean components in edible oil with ladder-shape melting temperature isothermal amplification (LMTIA) assay

Detection of Listeria monocytogenes in Food Using the Proofman-LMTIA Assay

Comparison of Plant Genomic DNA Extraction Kits Using the Proofman‐LMTIA Amplification Assay

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