Evaluation of species-specific PCR primers for detecting pork DNA in food seasoning products
Food adulteration remains a critical issue, particularly in highly processed foods where DNA degradation reduces the effectiveness of species identification. This study evaluated the performance of three different lengths of species-specific primers (398, 288 & 149 bp) targeting the mitochondria...
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| Format: | Article |
| Language: | en |
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Penerbit Universiti Kebangsaan Malaysia
2025
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| Online Access: | http://journalarticle.ukm.my/26538/1/ML%2012.pdf http://journalarticle.ukm.my/26538/ https://jms.mabjournal.com/index.php/mab/issue/view/69 |
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| author | Nur Atiqah Khairul Adha, Lesley Maurice Bilung, Aida Azrina Azmi, Awang Ahmad Sallehin Awang Husaini, Zaliha Suadi, |
| author_facet | Nur Atiqah Khairul Adha, Lesley Maurice Bilung, Aida Azrina Azmi, Awang Ahmad Sallehin Awang Husaini, Zaliha Suadi, |
| author_sort | Nur Atiqah Khairul Adha, |
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| content_provider | Universiti Kebangsaan Malaysia |
| content_source | UKM Journal Article Repository |
| continent | Asia |
| country | Malaysia |
| description | Food adulteration remains a critical issue, particularly in highly processed foods where DNA degradation reduces the effectiveness of species identification. This study evaluated the performance of three different lengths of species-specific primers (398, 288 & 149 bp) targeting the mitochondrial cytochrome B gene in pork and aimed to determine the most effective primer for detecting pork DNA in food seasoning products using species-specific Polymerase Chain Reaction (PCR). Furthermore, species-specific primers for bovine and chicken were applied as controls to confirm species identification. The DNA was extracted from raw meat, binary mixtures, and seasonings. Sensitivity was tested with low pork concentrations, and the applicability of pork-specific primers was further evaluated in seasoning products without halal certification to assess potential pork adulteration. The results show that the DNA extracted from raw meat samples exhibited high purity (OD260/280:1.82 –2.00) and concentration (114.83 to 257.16 ng/µL), whereas food seasoning products yielded extremely lower values of purity (OD260/280:0.97 – 1.81) and concentration (2.75 to 66.18 ng/µL). Despite DNA degradation in processed foods, PCR products remain detectable. The shortest 149 bp primer demonstrated the highest sensitivity among the primers, successfully detecting pork DNA at a minimum concentration of 1% (w/w) in a binary meat mixture. The results demonstrated the absence of pork DNA in all non-pork labelled samples. However, the detection of chicken DNA in fermented pork cube samples indicated potential mislabelling or cross contamination during processing. These findings emphasise the importance of primer selection for detecting highly degraded DNA. This also underscores the applicability of species-specific PCR as a practical and robust approach for routine food authentication, providing a valuable tool to ensure halal compliance within the food industry. |
| format | Article |
| id | my-ukm.journal.26538 |
| institution | Universiti Kebangsaan Malaysia |
| language | en |
| publishDate | 2025 |
| publisher | Penerbit Universiti Kebangsaan Malaysia |
| record_format | eprints |
| spelling | my-ukm.journal.265382026-02-06T01:10:34Z http://journalarticle.ukm.my/26538/ Evaluation of species-specific PCR primers for detecting pork DNA in food seasoning products Nur Atiqah Khairul Adha, Lesley Maurice Bilung, Aida Azrina Azmi, Awang Ahmad Sallehin Awang Husaini, Zaliha Suadi, Food adulteration remains a critical issue, particularly in highly processed foods where DNA degradation reduces the effectiveness of species identification. This study evaluated the performance of three different lengths of species-specific primers (398, 288 & 149 bp) targeting the mitochondrial cytochrome B gene in pork and aimed to determine the most effective primer for detecting pork DNA in food seasoning products using species-specific Polymerase Chain Reaction (PCR). Furthermore, species-specific primers for bovine and chicken were applied as controls to confirm species identification. The DNA was extracted from raw meat, binary mixtures, and seasonings. Sensitivity was tested with low pork concentrations, and the applicability of pork-specific primers was further evaluated in seasoning products without halal certification to assess potential pork adulteration. The results show that the DNA extracted from raw meat samples exhibited high purity (OD260/280:1.82 –2.00) and concentration (114.83 to 257.16 ng/µL), whereas food seasoning products yielded extremely lower values of purity (OD260/280:0.97 – 1.81) and concentration (2.75 to 66.18 ng/µL). Despite DNA degradation in processed foods, PCR products remain detectable. The shortest 149 bp primer demonstrated the highest sensitivity among the primers, successfully detecting pork DNA at a minimum concentration of 1% (w/w) in a binary meat mixture. The results demonstrated the absence of pork DNA in all non-pork labelled samples. However, the detection of chicken DNA in fermented pork cube samples indicated potential mislabelling or cross contamination during processing. These findings emphasise the importance of primer selection for detecting highly degraded DNA. This also underscores the applicability of species-specific PCR as a practical and robust approach for routine food authentication, providing a valuable tool to ensure halal compliance within the food industry. Penerbit Universiti Kebangsaan Malaysia 2025 Article PeerReviewed application/pdf en http://journalarticle.ukm.my/26538/1/ML%2012.pdf Nur Atiqah Khairul Adha, and Lesley Maurice Bilung, and Aida Azrina Azmi, and Awang Ahmad Sallehin Awang Husaini, and Zaliha Suadi, (2025) Evaluation of species-specific PCR primers for detecting pork DNA in food seasoning products. Malaysian Applied Biology, 54 (4). pp. 130-138. ISSN 0126-8643 https://jms.mabjournal.com/index.php/mab/issue/view/69 |
| spellingShingle | Nur Atiqah Khairul Adha, Lesley Maurice Bilung, Aida Azrina Azmi, Awang Ahmad Sallehin Awang Husaini, Zaliha Suadi, Evaluation of species-specific PCR primers for detecting pork DNA in food seasoning products |
| title | Evaluation of species-specific PCR primers for detecting pork DNA in food seasoning products |
| title_full | Evaluation of species-specific PCR primers for detecting pork DNA in food seasoning products |
| title_fullStr | Evaluation of species-specific PCR primers for detecting pork DNA in food seasoning products |
| title_full_unstemmed | Evaluation of species-specific PCR primers for detecting pork DNA in food seasoning products |
| title_short | Evaluation of species-specific PCR primers for detecting pork DNA in food seasoning products |
| title_sort | evaluation of species-specific pcr primers for detecting pork dna in food seasoning products |
| url | http://journalarticle.ukm.my/26538/1/ML%2012.pdf http://journalarticle.ukm.my/26538/ https://jms.mabjournal.com/index.php/mab/issue/view/69 |
| url_provider | http://journalarticle.ukm.my/ |