Enhancing Food Transparency: PCR-HRM-Based Multiplex Assay for Simultaneous Detection of Four Animal Species in Food

To address the growing global issue of food fraud through animal species substitution, a quadruplex polymerase chain reaction combined with a high-resolution melting assay was developed in a single tube to rapidly identify the <i>cytb</i> DNA of four animal species. Custom primers were meticulously designed and experimentally validated; the <i>T</i><sub>m</sub> values for porcine, bovine, donkey, and tilapia amplicons were 72.28, 76.25, 79.32, and 84.14 °C, respectively. The results demonstrated that the method possessed high specificity (8:8), with a template DNA detection limit as low as 0.05 ng/μL, and was capable of reliably identifying 1% porcine adulteration in diverse food matrices. Moreover, this method was employed to detect animal-derived constituents within diverse processed food samples. The PCR-HRM assay was a rapid, cost-effective, and sensitive tool for food authenticity verification.

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Publication Details

Journal
Journal of Agricultural and Food Chemistry
Published
2025-12-05
DOI
https://doi.org/10.1021/acs.jafc.5c10963
Primary Topic
Identification and Quantification in Food
Type
article
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article

Enhancing Food Transparency: PCR-HRM-Based Multiplex Assay for Simultaneous Detection of Four Animal Species in Food

Shan Shan, Hao Luo, Wenyu Du, Yanmei Huang et al.
Journal of Agricultural and Food Chemistry
Identification and Quantification in Food
article

Enhancing Food Transparency: PCR-HRM-Based Multiplex Assay for Simultaneous Detection of Four Animal Species in Food

Shan Shan, Hao Luo, Wenyu Du, Yanmei Huang, Long Cheng, Yuanjian Zhang, Xiao‐Mei Sha, Zongcai Tu, J. L. Cheng, Haibin Liu, Daofeng Liu, Yang Luo, Wenming Xie
article en

Abstract

To address the growing global issue of food fraud through animal species substitution, a quadruplex polymerase chain reaction combined with a high-resolution melting assay was developed in a single tube to rapidly identify the <i>cytb</i> DNA of four animal species. Custom primers were meticulously designed and experimentally validated; the <i>T</i><sub>m</sub> values for porcine, bovine, donkey, and tilapia amplicons were 72.28, 76.25, 79.32, and 84.14 °C, respectively. The results demonstrated that the method possessed high specificity (8:8), with a template DNA detection limit as low as 0.05 ng/μL, and was capable of reliably identifying 1% porcine adulteration in diverse food matrices. Moreover, this method was employed to detect animal-derived constituents within diverse processed food samples. The PCR-HRM assay was a rapid, cost-effective, and sensitive tool for food authenticity verification.

Journal of Agricultural and Food ChemistryVol. 73(50)
Jiujiang University (CN), Liaocheng University (CN), Henan Tianguan Group (China) (CN), Beike Biotechnology (China) (CN), ID-FISH Technology (United States) (US), Bio-Medical Science (South Korea) (KR), Dà-Jiāng Innovations Science and Technology (China) (CN), Dong-A Pharmaceutical (South Korea) (KR), Nanchang Center for Disease Control and Prevention (CN), Food Research and Development Institute (BG), Jiangxi Normal University (CN)
Openalex Percentile: Top 65%
Identification and Quantification in Food
DOISourceOpenAlex
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