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Species identification and quantification of silver pomfret using the droplet digital PCR assay
- Source :
- Food Chemistry. 302:125331
- Publication Year :
- 2020
- Publisher :
- Elsevier BV, 2020.
-
Abstract
- Adulteration of the high-value silver pomfret (Pampus argenteus) is a serious problem worldwide, necessitating accurate identification and quantification of the species. In this study, optimisation of the digital droplet PCR (ddPCR) assay for the identification and quantification of the silver pomfret was carried out. The primer and probe concentrations, melting temperature, and PCR cycle number were optimised by combining single-factor experiments with an orthogonal experimental design. The absolute limits of detection and quantification of the ddPCR were 2copies/μl and 21 copies/μl, respectively. Its sensitivity was 0.1% for meat mixtures and 0.5% for DNA mixtures. The ddPCR was 156 times more sensitive than the real-time PCR, although both methods had similar specificities. However, the overall time needed to complete the ddPCR method was twice that of the real-time PCR. Notwithstanding, the ddPCR methodology established in this study can be a valuable tool for addressing species adulteration issues.
- Subjects :
- Melting temperature
Polymerase Chain Reaction
Sensitivity and Specificity
01 natural sciences
Analytical Chemistry
law.invention
0404 agricultural biotechnology
law
Fish Products
Animals
Species identification
Digital polymerase chain reaction
Digital droplet pcr
Polymerase chain reaction
Detection limit
Pomfret
Chromatography
biology
Chemistry
010401 analytical chemistry
Reproducibility of Results
04 agricultural and veterinary sciences
General Medicine
biology.organism_classification
040401 food science
Perciformes
0104 chemical sciences
Food Analysis
Food Science
Subjects
Details
- ISSN :
- 03088146
- Volume :
- 302
- Database :
- OpenAIRE
- Journal :
- Food Chemistry
- Accession number :
- edsair.doi.dedup.....20c469e0ad68626a164c7765a611d6bf