Development of a Real-Time PCR Assay for the Detection of Donkey (Equus asinus) Meat in Meat Mixtures Treated under Different Processing Conditions

In this study, a donkey-specific primer pair and probe were designed from mitochondrial cytochrome b gene for the detection of raw donkey meat and different processed meat mixtures. The PCR product size for donkey DNA was 99 bp, and primer specificity was verified using 20 animal species. The limit...

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Bibliographic Details
Published in:Foods Vol. 9; no. 2; p. 130
Main Authors: Kim, Mi-Ju, Suh, Seung-Man, Kim, Sung-Yeon, Qin, Pei, Kim, Hong-Rae, Kim, Hae-Yeong
Format: Journal Article
Language:English
Published: Switzerland MDPI 26.01.2020
MDPI AG
Subjects:
asses
autoclaving
beef
cytochrome b
detection limit
DNA
donkey
Equus asinus
food adulteration
food fraud
genes
meat products
mitochondria
oligodeoxyribonucleotides
processed meat
quantitative polymerase chain reaction
real-time pcr
donkey
meat products
real-time PCR
food adulteration
food fraud
cytochrome b
ISSN:2304-8158, 2304-8158
Online Access:Get full text
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Summary:In this study, a donkey-specific primer pair and probe were designed from mitochondrial cytochrome b gene for the detection of raw donkey meat and different processed meat mixtures. The PCR product size for donkey DNA was 99 bp, and primer specificity was verified using 20 animal species. The limit of detection (LOD) was examined by serially diluting donkey DNA. Using real-time PCR, 0.001 ng of donkey DNA could be detected. In addition, binary meat mixtures with various percentages of donkey meat (0.001%, 0.01%, 0.1%, 1%, 10%, and 100%) in beef were analyzed to determine the sensitivity of this real-time PCR assay. At least 0.001% of donkey meat was detected in raw, boiled, roasted, dried, grinded, fried, and autoclaved meat mixtures. The developed real-time PCR method showed sufficient specificity and sensitivity in identification of donkey meat and could be a useful tool for the identification of donkey meat in processed products.
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These authors contributed equally to the work.
ISSN:2304-8158
2304-8158
DOI:10.3390/foods9020130