Application of real-time PCR for analysis canine meat (Canis lupus familiaris) in goat's satay for halal authentication study
Background: Canine meat (CM) is one of the non-halal meats prohibited for consumption by the Muslim community. Due to low price compared with beef, CM is typically used as meat adulterants in halal-food based products such as Satay and meatballs to get the economic profits. Aim: The objective of...
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| Format: | Article |
| Language: | English |
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Tripoli University
2025-01-01
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| Series: | Open Veterinary Journal |
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| Online Access: | http://www.ejmanager.com/fulltextpdf.php?mno=220693 |
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| author | Lily Arsanti Lestari Wan Nurul Aini Marlyn Dian Laksitorini Yuny Erwanto Agustina Ari Murti Budi Hastuti Mohammad Zainal Abidin Nurulia Hidayah Muhammad Jefriyanto Budikafa Abdul Rohman |
| author_facet | Lily Arsanti Lestari Wan Nurul Aini Marlyn Dian Laksitorini Yuny Erwanto Agustina Ari Murti Budi Hastuti Mohammad Zainal Abidin Nurulia Hidayah Muhammad Jefriyanto Budikafa Abdul Rohman |
| author_sort | Lily Arsanti Lestari |
| collection | DOAJ |
| description | Background:
Canine meat (CM) is one of the non-halal meats prohibited for consumption by the Muslim community. Due to low price compared with beef, CM is typically used as meat adulterants in halal-food based products such as Satay and meatballs to get the economic profits.
Aim:
The objective of this study was to design a novel species-specific primer in combination with real-time polymerase chain reaction for analysis of Canine's DNA for halal authentication analysis.
Methods:
A Primer targeting the D-loop region of mitochondrial DNA was designed and subjected to validation procedure by assessing some performance characteristics including specificity, amplification efficiency (E), sensitivity, repeatability and linearity describing the correlation between concentration of Canine's DNA (x-axis) and quantification cycle (Cq) in y-axis. The designed primer was specific over other meat DNAs applying the annealing temperature (Tm) of 57.8 °C.
Results:
RT-PCR method produced an acceptable amplification efficiency (E) of 109.7% with the coefficient of determination (R2) for the correlation between Cq and log DNA concentration of 0.999. The sensitivity of developed method provides limit of detection (LOD) value of 31.25 pg/µL. The precision of analytical method is acceptable with relative standard deviation (RSD) value of 2%. The method with designed D-loop primer was successfully applied for detection and quantification of Canine's DNA in food products. There are no amplification profiles for Canine DNA in marketed goat's satay products.
Conclusion:
RT-PCR combined with novel primer targeting D-loop provide a specific and accurate analytical tool for identification of CM for halal authentication studies. [Open Vet J 2025; 15(1.000): 456-464] |
| format | Article |
| id | doaj-art-ceac33e5a0434438b47ade132ae292ce |
| institution | OA Journals |
| issn | 2226-4485 2218-6050 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Tripoli University |
| record_format | Article |
| series | Open Veterinary Journal |
| spelling | doaj-art-ceac33e5a0434438b47ade132ae292ce2025-08-20T02:16:05ZengTripoli UniversityOpen Veterinary Journal2226-44852218-60502025-01-0115145646410.5455/OVJ.2025.v15.i1.41220693Application of real-time PCR for analysis canine meat (Canis lupus familiaris) in goat's satay for halal authentication studyLily Arsanti Lestari0Wan Nurul Aini1Marlyn Dian Laksitorini2Yuny Erwanto3Agustina Ari Murti Budi Hastuti4Mohammad Zainal Abidin5Nurulia Hidayah6Muhammad Jefriyanto Budikafa7Abdul Rohman8Department of Nutrition and Health, Faculty of Medicine, Public Health and Nursing, Universitas Gadjah Mada, Yogyakarta 55281, Indonesia. Faculty of Pharmacy Universitas Gadjah Mada, Yogyakarta 55281, Indonesia. Faculty of Pharmacy Universitas Gadjah Mada, Yogyakarta 55281, Indonesia Center of Excellence, Institute for Halal Industry and System (IHIS), Universitas Gadjah Mada, Yogyakarta 55281, Indonesia. Faculty of Pharmacy Universitas Gadjah Mada, Yogyakarta 55281, Indonesia. Faculty of Animal Science, Universitas Gadjah Mada, Yogyakarta 55281, Indonesia. Field Research Center, Universitas Gadjah Mada, Yogyakarta 55281, Indonesia Faculty of Islamic Economics and Business, State Islamic University Prof. K. H. Saifuddin Zuhri, Purwokerto 53126, Indonesia. Center of Excellence, Institute for Halal Industry and System (IHIS), Universitas Gadjah Mada, Yogyakarta 55281, Indonesia.Background: Canine meat (CM) is one of the non-halal meats prohibited for consumption by the Muslim community. Due to low price compared with beef, CM is typically used as meat adulterants in halal-food based products such as Satay and meatballs to get the economic profits. Aim: The objective of this study was to design a novel species-specific primer in combination with real-time polymerase chain reaction for analysis of Canine's DNA for halal authentication analysis. Methods: A Primer targeting the D-loop region of mitochondrial DNA was designed and subjected to validation procedure by assessing some performance characteristics including specificity, amplification efficiency (E), sensitivity, repeatability and linearity describing the correlation between concentration of Canine's DNA (x-axis) and quantification cycle (Cq) in y-axis. The designed primer was specific over other meat DNAs applying the annealing temperature (Tm) of 57.8 °C. Results: RT-PCR method produced an acceptable amplification efficiency (E) of 109.7% with the coefficient of determination (R2) for the correlation between Cq and log DNA concentration of 0.999. The sensitivity of developed method provides limit of detection (LOD) value of 31.25 pg/µL. The precision of analytical method is acceptable with relative standard deviation (RSD) value of 2%. The method with designed D-loop primer was successfully applied for detection and quantification of Canine's DNA in food products. There are no amplification profiles for Canine DNA in marketed goat's satay products. Conclusion: RT-PCR combined with novel primer targeting D-loop provide a specific and accurate analytical tool for identification of CM for halal authentication studies. [Open Vet J 2025; 15(1.000): 456-464]http://www.ejmanager.com/fulltextpdf.php?mno=220693d-loop primerreal time-pcrcanine meatgoat's satay |
| spellingShingle | Lily Arsanti Lestari Wan Nurul Aini Marlyn Dian Laksitorini Yuny Erwanto Agustina Ari Murti Budi Hastuti Mohammad Zainal Abidin Nurulia Hidayah Muhammad Jefriyanto Budikafa Abdul Rohman Application of real-time PCR for analysis canine meat (Canis lupus familiaris) in goat's satay for halal authentication study Open Veterinary Journal d-loop primer real time-pcr canine meat goat's satay |
| title | Application of real-time PCR for analysis canine meat (Canis lupus familiaris) in goat's satay for halal authentication study |
| title_full | Application of real-time PCR for analysis canine meat (Canis lupus familiaris) in goat's satay for halal authentication study |
| title_fullStr | Application of real-time PCR for analysis canine meat (Canis lupus familiaris) in goat's satay for halal authentication study |
| title_full_unstemmed | Application of real-time PCR for analysis canine meat (Canis lupus familiaris) in goat's satay for halal authentication study |
| title_short | Application of real-time PCR for analysis canine meat (Canis lupus familiaris) in goat's satay for halal authentication study |
| title_sort | application of real time pcr for analysis canine meat canis lupus familiaris in goat apos s satay for halal authentication study |
| topic | d-loop primer real time-pcr canine meat goat's satay |
| url | http://www.ejmanager.com/fulltextpdf.php?mno=220693 |
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