Comparison of PCR Techniques in Adulteration Identification of Dairy Products
Abstract
:1. Introduction
2. DNA Extraction Methods
2.1. Organic Solvent Extraction Method
2.2. Salting-Out Extraction Method
2.3. Silicon Centrifugal Column Extraction Method
2.4. Magnetic Bead Extraction Method
3. PCR-Based Dairy Product Authenticity Testing
3.1. PCR-RFLP
3.2. Multiplex PCR
3.3. Real-Time PCR
3.4. Digital PCR (dPCR)
3.5. Randomly Amplified Polymorphic DNA PCR (RAPD-PCR)
4. Factors Affecting PCR-Based Dairy Product Authenticity Testing
4.1. Disruption of DNA Integrity by Food Processing
4.2. PCR Inhibitors
5. Conclusions and Future Perspectives
Author Contributions
Funding
Institutional Review Board Statement
Data Availability Statement
Conflicts of Interest
References
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Extraction Method | Advantages | Disadvantages | Refs. |
---|---|---|---|
Organic solvent extraction method | High purity; Large quantity; Intact fragments; Low cost | Use of toxic reagents; Poor reproducibility; Time-consuming | [25,26,35] |
Salting-out extraction method | Simplicity; No use of toxic reagents; Highly efficient | Low yield; Less application range; Large quantities of raw material | [12,28,29] |
Silicon centrifugal columns extraction method | Higher integrity and purity; Easy operation; Less time consumption | More pronounced shearing of genomic DNA; Higher cost; Lower concentration of DNA | [28,36,37] |
Magnetic beads extraction method | Less time consumption; Easy operation; Efficient | Low yield | [33,34,36] |
Techniques | Species Identification | Types of Dairy Products | DNA Target | Detection Limit | Ref. |
---|---|---|---|---|---|
PCR-RFLP | Identification of the milk sources from cow, sheep, and buffalo milk | Raw milk | cyt b gene | Qualitative analysis | [42] |
Identification of the milk sources from cow, sheep, goat, and buffalo | Raw milk | cyt b gene | Qualitative analysis | [43] | |
Identification of the milk sources from cow, sheep, and buffalo milk | Raw milk | κ-casein gene | Qualitative analysis | [44] | |
Identification of the milk sources from three Egyptian goat breeds (Bariki, Damascus, and Zaraibl) | Blood sample | myostatin (MSTN) gene and prolactin (PRL) gene | Qualitative analysis | [74] | |
Multiple PCR | Identification of the cheese sources from cow, sheep, and goat milk | Cheese | Mitochondrial 12S and 16S rRNA genes. | 0.5% detection limit | [45] |
Identification of the milk sources from cow, goat, and sheep milk | Raw milk | cyt b genes (cow and sheep) D-loop range (goats) | 0.1% detection limit. | [46] | |
Identification of the milk sources from camel, horse, and goat | Raw milk | 16S rRNA gene and D-LOOP range | 0.1% detection limit. | [50] | |
Identification of the milk sources from cow, goat, sheep, and buffalo milk | Raw milk | cyt b gene and 12S rRNA | 0.1% detection limit. | [55] | |
Identification of the milk sources from cow, sheep, and goat milk | Blood sample | 16S rRNA (sheep) and 12S RNA (cow, sheep, and goat) | 0.1% detection limit. | [75] | |
Real-time TaqMan PCR | Identification of goat milk or cow milk from sheep milk | Raw milk | 12S rRNA | Linear dynamic range of 0.6–10% | [48] |
Identification of the milk sources from cow and mare milk | Raw milk | 12S rRNA | 1 pg of cow DNA; 1 pg of mare DNA, | [51] | |
Identification of the cheese, and milk sources from cow and goat milk | Cheese and raw milk | 12S rRNA | 0.005 ng/μL in milk and 0.01 ng/μL in cheese | [52] | |
Identification of the cheese and milk sources of sheep and goat milk | Cheese and raw milk | 12S rRNA | 0.005 ng/μL in milk and 0.01 ng/μL in cheese | [53] | |
Identification of the milk, yogurt, cheese, milk powder, and milk beverage sources from cow and camel milk | Milk, yogurt, cheese, milk powder, and milk beverage | 12S rRNA | 0.0025 to 0.001 ng/μL (milk); 0.5 to 0.001 ng/μL (yogurt); 1 to 0.05 ng/μL (cheese); 0.01 ng/μL (milk powder); 0.001 ng/μL (milk beverage) | [54] | |
Real-time SYBR Green PCR | Identify the cheese source from cow and buffalo milk | Cheese | Cytochrome oxidase subunit 1 (coI) | - a | [47] |
Identification of the milk sources from cow, and buffalo | Cheese | Cytochrome oxidase subunit 1 (coI) | 0.5 ng/μL | [76] | |
Identification of the milk sources from camel, cow, and goat | Milk, milk powder, and milk soap | Follicle stimulating hormone receptor (FSHR) gene (camel and goat) and 12S RNA (cow) | Rang of 0.001–0.002% | [77] | |
Digital PCR | Identify the cheese source from cow and buffalo milk | Cheese | cyt b genes | 0.1% detection limit. | [64] |
RAPD-PCR | Identify the cheese source from four sheep breeds’ milk | Cheese | Sequence-characterized amplified region | Qualitative analysis | [73] |
PCR Techniques | Cost | Detect Time | Analysis Procedure | Other |
---|---|---|---|---|
PCR-RFLP | Low | Fast | Not quantitative analysis. | Simple. |
Multiple PCR | Low | Fast | Both qualitative and quantitative analyses. | High-throughput; False-positive result |
Real-time PCR | Low | Fast | Both qualitative and quantitative analyses. | Excellent sensitivity; Specificity; High consistency; False-positive result |
Digital PCR | High | Fast | Both qualitative and quantitative analyses. | Accurate; Highly reproducible; Narrow dynamic range |
RAPD-PCR | Low | Fast | Not quantitative analysis. | Independent from Specific primers; Ease to use; Less interfering factors |
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Li, B.; Yu, M.; Xu, W.; Chen, L.; Han, J. Comparison of PCR Techniques in Adulteration Identification of Dairy Products. Agriculture 2023, 13, 1450. https://doi.org/10.3390/agriculture13071450
Li B, Yu M, Xu W, Chen L, Han J. Comparison of PCR Techniques in Adulteration Identification of Dairy Products. Agriculture. 2023; 13(7):1450. https://doi.org/10.3390/agriculture13071450
Chicago/Turabian StyleLi, Baiyi, Mingxue Yu, Weiping Xu, Lu Chen, and Juan Han. 2023. "Comparison of PCR Techniques in Adulteration Identification of Dairy Products" Agriculture 13, no. 7: 1450. https://doi.org/10.3390/agriculture13071450