A Rapid RPA-CRISPR/Cas12a Detection Method for Adulteration of Goat Milk Powder
Because of the serious adulteration of goat milk, the rapid on-site detection of goat milk powder adulteration is needed. In this study, the CRISPR/Cas12a detection system combined with recombinase polymerase amplification (RPA) was employed to qualitatively detect the adulteration of goat milk powd...
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| Format: | Article |
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MDPI AG
2023-04-01
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| Series: | Foods |
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| Online Access: | https://www.mdpi.com/2304-8158/12/8/1569 |
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| author | Shuqin Huang Yan Liu Xu Zhang Zuoqi Gai Hongtao Lei Xing Shen |
| author_facet | Shuqin Huang Yan Liu Xu Zhang Zuoqi Gai Hongtao Lei Xing Shen |
| author_sort | Shuqin Huang |
| collection | DOAJ |
| description | Because of the serious adulteration of goat milk, the rapid on-site detection of goat milk powder adulteration is needed. In this study, the CRISPR/Cas12a detection system combined with recombinase polymerase amplification (RPA) was employed to qualitatively detect the adulteration of goat milk powder with cattle-derived components. Specific primers and crRNA were designed and screened. After the optimization of RPA and the Cas system, the RPA-CRISPR/Cas12a detection method was established. The detection can complete the rapid identification of cattle-derived components in 45 min, without the assistant of large equipment. The absolute detectability of the RPA-CRISPR/Cas12a assay could reach 10<sup>−2</sup> ng/μL for cattle genomic DNA, and 1% (<i>w/w</i>) for cattle milk powder, which is suitable to meet the testing requirements for on-site detection. In total, 55 commercial goat milk powder products were collected for blind testing. The results showed that 27.3% of the samples were adulterated with cattle ingredients, revealing a serious adulteration situation in goat milk powder market. The RPA-CRISPR/Cas12a assay established in this research exhibited its potential for practical use of on-site detection to detect cow milk powder in goat milk powder and can provide reliable technical reference for combating food fraud of adulteration of goat milk products. |
| first_indexed | 2024-03-11T05:02:18Z |
| format | Article |
| id | doaj.art-9c03e769414c4efd90827f31d0fe4187 |
| institution | Directory Open Access Journal |
| issn | 2304-8158 |
| language | English |
| last_indexed | 2024-03-11T05:02:18Z |
| publishDate | 2023-04-01 |
| publisher | MDPI AG |
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| series | Foods |
| spelling | doaj.art-9c03e769414c4efd90827f31d0fe41872023-11-17T19:13:31ZengMDPI AGFoods2304-81582023-04-01128156910.3390/foods12081569A Rapid RPA-CRISPR/Cas12a Detection Method for Adulteration of Goat Milk PowderShuqin Huang0Yan Liu1Xu Zhang2Zuoqi Gai3Hongtao Lei4Xing Shen5Guangdong Provincial Key Laboratory of Food Quality and Safety/National-Local Joint Engineering Research Center for Machining and Safety of Livestock and Poultry Products, South China Agricultural University, Guangzhou 510642, ChinaGuangdong Provincial Key Laboratory of Food Quality and Safety/National-Local Joint Engineering Research Center for Machining and Safety of Livestock and Poultry Products, South China Agricultural University, Guangzhou 510642, ChinaGuangzhou Editgene Co., Ltd., Guangzhou 510630, ChinaGuangzhou Editgene Co., Ltd., Guangzhou 510630, ChinaGuangdong Provincial Key Laboratory of Food Quality and Safety/National-Local Joint Engineering Research Center for Machining and Safety of Livestock and Poultry Products, South China Agricultural University, Guangzhou 510642, ChinaGuangdong Provincial Key Laboratory of Food Quality and Safety/National-Local Joint Engineering Research Center for Machining and Safety of Livestock and Poultry Products, South China Agricultural University, Guangzhou 510642, ChinaBecause of the serious adulteration of goat milk, the rapid on-site detection of goat milk powder adulteration is needed. In this study, the CRISPR/Cas12a detection system combined with recombinase polymerase amplification (RPA) was employed to qualitatively detect the adulteration of goat milk powder with cattle-derived components. Specific primers and crRNA were designed and screened. After the optimization of RPA and the Cas system, the RPA-CRISPR/Cas12a detection method was established. The detection can complete the rapid identification of cattle-derived components in 45 min, without the assistant of large equipment. The absolute detectability of the RPA-CRISPR/Cas12a assay could reach 10<sup>−2</sup> ng/μL for cattle genomic DNA, and 1% (<i>w/w</i>) for cattle milk powder, which is suitable to meet the testing requirements for on-site detection. In total, 55 commercial goat milk powder products were collected for blind testing. The results showed that 27.3% of the samples were adulterated with cattle ingredients, revealing a serious adulteration situation in goat milk powder market. The RPA-CRISPR/Cas12a assay established in this research exhibited its potential for practical use of on-site detection to detect cow milk powder in goat milk powder and can provide reliable technical reference for combating food fraud of adulteration of goat milk products.https://www.mdpi.com/2304-8158/12/8/1569goat milk powderCRISPR/Cas12aadulterationrapid detectionfood fraud |
| spellingShingle | Shuqin Huang Yan Liu Xu Zhang Zuoqi Gai Hongtao Lei Xing Shen A Rapid RPA-CRISPR/Cas12a Detection Method for Adulteration of Goat Milk Powder Foods goat milk powder CRISPR/Cas12a adulteration rapid detection food fraud |
| title | A Rapid RPA-CRISPR/Cas12a Detection Method for Adulteration of Goat Milk Powder |
| title_full | A Rapid RPA-CRISPR/Cas12a Detection Method for Adulteration of Goat Milk Powder |
| title_fullStr | A Rapid RPA-CRISPR/Cas12a Detection Method for Adulteration of Goat Milk Powder |
| title_full_unstemmed | A Rapid RPA-CRISPR/Cas12a Detection Method for Adulteration of Goat Milk Powder |
| title_short | A Rapid RPA-CRISPR/Cas12a Detection Method for Adulteration of Goat Milk Powder |
| title_sort | rapid rpa crispr cas12a detection method for adulteration of goat milk powder |
| topic | goat milk powder CRISPR/Cas12a adulteration rapid detection food fraud |
| url | https://www.mdpi.com/2304-8158/12/8/1569 |
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