A high sensitivity method of closed-tube loop-mediated isothermal amplification developed for visual and rapid detection of cow milk adulteration

Wang, Yifan; Yu, Zhezhe; Liu, Yongfeng

doi:10.1016/j.idairyj.2021.105214

Cited by 8 publications

(1 citation statement)

References 39 publications

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“…At present, there are many milk protein allergen detection methods, which can be divided into two categories in terms of detection objects. One category chooses DNA as the object, such as polymerase chain reaction (PCR) and loop-mediated isothermal amplification (LAMP) [ 4 , 5 , 6 , 7 ]. However, since these methods detect the non-allergenic DNA but not the allergen protein itself, it may be independent of the amount of protein in an unknown substrate.…”

Section: Introductionmentioning

confidence: 99%

Identification of Characteristic Peptides of Casein in Cow Milk Based on MALDI-TOF MS for Direct Adulteration Detection of Goat Milk

Dai

Zhang

et al. 2023

Foods

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It is widely acknowledged that casein is an important allergenic protein in milk which may cause danger to customers. The identification and confirmation of caseins through mass spectrometry requires the selection of suitable characteristic peptides. In this study, by means of matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS), the three most representative specific peptides of caseins in cow milk were screened out with mass-to-charge ratios (m/z) of 830, 1195, and 1759, respectively. By comparing 2,5-dihydroxybenzoic acid (DHB) and α-cyano-4-hydroxycinnamic acid (CHCA) MALDI matrices, it was found that DHB was more suitable for peptide detection with the limits of detection (LODs) of 0.1 mg/L for α, β-casein. Furthermore, on the basis of verifying the characteristic peptides of casein from cow milk, this protocol was applied to goat milk authentication. Cow milk addition in goat milk was investigated by using the screened specific peptides. The results showed that the adulteration could be identified when the proportion of cow milk was 1% or more. When applied to inspect adulteration in five brands of commercial goat milk, specific peptides of bovine casein were detected in four of them. The method has the advantages of strong reliability, high throughput, simple preprocessing, and fast speed, which can provide powerful help for prewarning dairy allergen.

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Section: Introductionmentioning

confidence: 99%

Identification of Characteristic Peptides of Casein in Cow Milk Based on MALDI-TOF MS for Direct Adulteration Detection of Goat Milk

Dai

Zhang

et al. 2023

Foods

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Rapid DNA extraction and colorimetric amplicon visualisation speed up LAMP-based detection of soybean allergen in foods

Schäfer,

Allgöwer,

Holzhauser

2023

Eur Food Res Technol

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Detection of allergens in foods, including soybean, is relevant for food labelling requirements. Moreover, allergen-specific methods may allow standardisation of allergens in food matrices for use in food challenges as allergy diagnostic approaches. Rapid methods are preferred for screening and along the manufacturing line. Previously, we demonstrated sensitive and specific detection of soybean DNA by combining loop-mediated isothermal amplification (LAMP) and lateral flow device (LFD)-like visualisation. However, lengthy DNA extraction and potential contamination of subsequent by previous LAMP reactions from unclosed LFD may impact its use as a rapid and robust method. Here, we developed a rapid protocol for DNA extraction. Moreover, we identified phenol red for distinct visualisation of positive reactions in permanently closed reaction tubes. The optimised method was validated using complex foods (boiled sausage, instant soup, and chocolate) with known amounts of soybean. Further, its applicability was shown in 12 processed retail foods. Results were verified by orthogonal qPCR. The enhanced LAMP method allowed detection at or below 10 mg soybean per kg processed food. The method provides rapid and easy-to-use screening without the need for detection equipment. Hence, it may serve to verify the presence of soybean ingredients and support a risk-based precautionary labelling of non-ingredient soybean in compound foods. Also, as determination of clinical reaction thresholds before and after allergen immunotherapy (AIT) is both inclusion and exclusion criterion for clinical trials and success parameter of AIT, the method may allow verification of calculable soybean content in provocation meals and thus a standardised administration for threshold determination before and after AIT.

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Loop-Mediated Isothermal Amplification for On-Site Visual Identification of Leech Species

Peng,

Li,

Deng

et al. 2024

Food Anal. Methods

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A high sensitivity method of closed-tube loop-mediated isothermal amplification developed for visual and rapid detection of cow milk adulteration

Cited by 8 publications

References 39 publications

Identification of Characteristic Peptides of Casein in Cow Milk Based on MALDI-TOF MS for Direct Adulteration Detection of Goat Milk

Identification of Characteristic Peptides of Casein in Cow Milk Based on MALDI-TOF MS for Direct Adulteration Detection of Goat Milk

Rapid DNA extraction and colorimetric amplicon visualisation speed up LAMP-based detection of soybean allergen in foods

Loop-Mediated Isothermal Amplification for On-Site Visual Identification of Leech Species

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