Quantification of Pork, Chicken, Beef, and Sheep Contents in Meat Products Using Duplex Real-Time PCR
Accurate methods for meat speciation and quantification are essential for ensuring the supply of safe and wholesome meat and composite products with animal origins to negate the potential associated hazards, aid classification of consignments at the import control system, and thwart food fraud commi...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-08-01
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| Series: | Foods |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2304-8158/12/15/2971 |
| _version_ | 1827731209285795840 |
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| author | Yanwen Wang Emily Teo Kung Ju Lin Yuansheng Wu Joanne Sheot Harn Chan Li Kiang Tan |
| author_facet | Yanwen Wang Emily Teo Kung Ju Lin Yuansheng Wu Joanne Sheot Harn Chan Li Kiang Tan |
| author_sort | Yanwen Wang |
| collection | DOAJ |
| description | Accurate methods for meat speciation and quantification are essential for ensuring the supply of safe and wholesome meat and composite products with animal origins to negate the potential associated hazards, aid classification of consignments at the import control system, and thwart food fraud committed for financial gain. To better enhance meat safety control and combat food fraud, this study developed two duplex real-time polymerase chain reaction (real-time PCR) systems specifically designed for chicken, pork, sheep, and beef, using single-copy, chromosomally encoded, species-specific gene sequences to accurately measure the content of each meat type in meat products. DNA extracted from the raw and boiled reference materials prepared in varying proportions (ranging from 1% to 75%) were used in the development of the duplex assay to derive calibration factors to determine the meat content in different meat products. The method was further validated using proficiency test samples and market monitoring samples. Our findings showed that this method exhibits high specificity and sensitivity, with a significant accuracy range of 0.14% to 24.07% in quantifying the four meat types in both raw and processed meat products. Validation results further confirmed the effectiveness of our method in accurately quantifying meat content. Thus, we have demonstrated the duplex qPCR assays as promising approaches for implementation in routine analysis to strengthen meat safety control systems and combat meat fraud, thereby safeguarding consumer health and trust in the meat industry. |
| first_indexed | 2024-03-11T00:27:12Z |
| format | Article |
| id | doaj.art-4248029948bb4fe0a4b8dc4e43180268 |
| institution | Directory Open Access Journal |
| issn | 2304-8158 |
| language | English |
| last_indexed | 2024-03-11T00:27:12Z |
| publishDate | 2023-08-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Foods |
| spelling | doaj.art-4248029948bb4fe0a4b8dc4e431802682023-11-18T22:55:42ZengMDPI AGFoods2304-81582023-08-011215297110.3390/foods12152971Quantification of Pork, Chicken, Beef, and Sheep Contents in Meat Products Using Duplex Real-Time PCRYanwen Wang0Emily Teo1Kung Ju Lin2Yuansheng Wu3Joanne Sheot Harn Chan4Li Kiang Tan5National Centre for Food Science, Singapore Food Agency, 7 International Business Park, Singapore 609919, SingaporeNational Centre for Food Science, Singapore Food Agency, 7 International Business Park, Singapore 609919, SingaporeNational Centre for Food Science, Singapore Food Agency, 7 International Business Park, Singapore 609919, SingaporeNational Centre for Food Science, Singapore Food Agency, 7 International Business Park, Singapore 609919, SingaporeNational Centre for Food Science, Singapore Food Agency, 7 International Business Park, Singapore 609919, SingaporeNational Centre for Food Science, Singapore Food Agency, 7 International Business Park, Singapore 609919, SingaporeAccurate methods for meat speciation and quantification are essential for ensuring the supply of safe and wholesome meat and composite products with animal origins to negate the potential associated hazards, aid classification of consignments at the import control system, and thwart food fraud committed for financial gain. To better enhance meat safety control and combat food fraud, this study developed two duplex real-time polymerase chain reaction (real-time PCR) systems specifically designed for chicken, pork, sheep, and beef, using single-copy, chromosomally encoded, species-specific gene sequences to accurately measure the content of each meat type in meat products. DNA extracted from the raw and boiled reference materials prepared in varying proportions (ranging from 1% to 75%) were used in the development of the duplex assay to derive calibration factors to determine the meat content in different meat products. The method was further validated using proficiency test samples and market monitoring samples. Our findings showed that this method exhibits high specificity and sensitivity, with a significant accuracy range of 0.14% to 24.07% in quantifying the four meat types in both raw and processed meat products. Validation results further confirmed the effectiveness of our method in accurately quantifying meat content. Thus, we have demonstrated the duplex qPCR assays as promising approaches for implementation in routine analysis to strengthen meat safety control systems and combat meat fraud, thereby safeguarding consumer health and trust in the meat industry.https://www.mdpi.com/2304-8158/12/15/2971meat safetyfood fraudmeat speciationmeat quantificationduplex qPCRchicken |
| spellingShingle | Yanwen Wang Emily Teo Kung Ju Lin Yuansheng Wu Joanne Sheot Harn Chan Li Kiang Tan Quantification of Pork, Chicken, Beef, and Sheep Contents in Meat Products Using Duplex Real-Time PCR Foods meat safety food fraud meat speciation meat quantification duplex qPCR chicken |
| title | Quantification of Pork, Chicken, Beef, and Sheep Contents in Meat Products Using Duplex Real-Time PCR |
| title_full | Quantification of Pork, Chicken, Beef, and Sheep Contents in Meat Products Using Duplex Real-Time PCR |
| title_fullStr | Quantification of Pork, Chicken, Beef, and Sheep Contents in Meat Products Using Duplex Real-Time PCR |
| title_full_unstemmed | Quantification of Pork, Chicken, Beef, and Sheep Contents in Meat Products Using Duplex Real-Time PCR |
| title_short | Quantification of Pork, Chicken, Beef, and Sheep Contents in Meat Products Using Duplex Real-Time PCR |
| title_sort | quantification of pork chicken beef and sheep contents in meat products using duplex real time pcr |
| topic | meat safety food fraud meat speciation meat quantification duplex qPCR chicken |
| url | https://www.mdpi.com/2304-8158/12/15/2971 |
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