The employment of q-PCR using specific primer targeting on mitochondrial cytochrome-b gene for identification of wild boar meat in meatball samples
Objective: The objective of this study was to employ real-time or quantitative polymerase chain reaction (q-PCR) using novel species specific primer (SSP) targeting on mitochondrial cytochrome-b of wild boar species (CYTBWB2-wb) gene for the identification of non-halal meat of wild boar meat (WBM) i...
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Network for the Veterinarians of Bangladesh
2019-09-01
|
| Series: | Journal of Advanced Veterinary and Animal Research |
| Subjects: | |
| Online Access: | http://www.ejmanager.com/fulltextpdf.php?mno=17408 |
| _version_ | 1818138967931879424 |
|---|---|
| author | Ganea Qorry Aina Yuny Erwanto Motalib Hossain Mohd Rafie Johan Md. Eaqub Ali Abdul Rohman |
| author_facet | Ganea Qorry Aina Yuny Erwanto Motalib Hossain Mohd Rafie Johan Md. Eaqub Ali Abdul Rohman |
| author_sort | Ganea Qorry Aina |
| collection | DOAJ |
| description | Objective: The objective of this study was to employ real-time or quantitative polymerase chain reaction (q-PCR) using novel species specific primer (SSP) targeting on mitochondrial cytochrome-b of wild boar species (CYTBWB2-wb) gene for the identification of non-halal meat of wild boar meat (WBM) in meatball products.
Materials and Methods: The novel SSP of CYTBWB2-wb was designed by our group using PRIMERQUEST and NCBI software. DNA was extracted using propanol-chloroform-isoamyl alcohol method. The designed SSP was further subjected for validation protocols using DNA isolated from fresh meat and from meatball, which include specificity test, determination of efficiency, limit of detection and repeatability, and application of developed method for analysis of commercially meatball samples
Results: The results showed that CYTBWB2-wb was specific to wild boar species against other animal species with optimized annealing temperature of 59°C. The efficiency of q-PCR obtained was 91.9% which is acceptable according to the Codex Allimentarius Commission (2010). DNA, with as low as 5 pg/μl, could be detected using q-PCR with primer of CYTBWB2-wb. The developed method was also used for DNA analysis extracted from meatball samples commercially available.
Conclusion: q-PCR using CYTBWB2-wb primers targeting on mitochondrial cytochrome-b gene (forward: CGG TTC CCT CTT AGG CAT TT; Reverse: GGA TGA ACA GGC AGA TGA AGA) can be fruitfully used for the analysis of WBM in commercial meatball samples. [J Adv Vet Anim Res 2019; 6(3.000): 300-307] |
| first_indexed | 2024-12-11T10:20:37Z |
| format | Article |
| id | doaj.art-cc5fb35b08a247dab496fa1f6e09f2d7 |
| institution | Directory Open Access Journal |
| issn | 2311-7710 |
| language | English |
| last_indexed | 2024-12-11T10:20:37Z |
| publishDate | 2019-09-01 |
| publisher | Network for the Veterinarians of Bangladesh |
| record_format | Article |
| series | Journal of Advanced Veterinary and Animal Research |
| spelling | doaj.art-cc5fb35b08a247dab496fa1f6e09f2d72022-12-22T01:11:28ZengNetwork for the Veterinarians of BangladeshJournal of Advanced Veterinary and Animal Research2311-77102019-09-016330030710.5455/javar.2019.f34817408The employment of q-PCR using specific primer targeting on mitochondrial cytochrome-b gene for identification of wild boar meat in meatball samplesGanea Qorry Aina0Yuny Erwanto1Motalib Hossain2Mohd Rafie Johan3Md. Eaqub Ali4Abdul Rohman5Department of Pharmaceutical Chemistry, Gadjah Mada University, Yogyakarta, Indonesia. & Department of Medical Laboratory Technology, Politeknik Kesehatan Kementerian Kesehatan Kalimantan Timur, Kalimantan Timur, Indonesia Division of Animal Products Technology, Faculty of Animal Science, Yogyakarta, Indonesia. & Research Centre of Halal Products, Gadjah Mada University, Yogyakarta, Indonesia Nanotechnology and Catalysis Research Centre (NanoCat), University of Malaya, Kuala Lumpur, Malaysia Nanotechnology and Catalysis Research Centre (NanoCat), University of Malaya, Kuala Lumpur, Malaysia Nanotechnology and Catalysis Research Centre (NanoCat), University of Malaya, Kuala Lumpur, Malaysia Department of Pharmaceutical Chemistry, Gadjah Mada University, Yogyakarta, Indonesia. & Research Centre of Halal Products, Gadjah Mada University, Yogyakarta, Indonesia.Objective: The objective of this study was to employ real-time or quantitative polymerase chain reaction (q-PCR) using novel species specific primer (SSP) targeting on mitochondrial cytochrome-b of wild boar species (CYTBWB2-wb) gene for the identification of non-halal meat of wild boar meat (WBM) in meatball products. Materials and Methods: The novel SSP of CYTBWB2-wb was designed by our group using PRIMERQUEST and NCBI software. DNA was extracted using propanol-chloroform-isoamyl alcohol method. The designed SSP was further subjected for validation protocols using DNA isolated from fresh meat and from meatball, which include specificity test, determination of efficiency, limit of detection and repeatability, and application of developed method for analysis of commercially meatball samples Results: The results showed that CYTBWB2-wb was specific to wild boar species against other animal species with optimized annealing temperature of 59°C. The efficiency of q-PCR obtained was 91.9% which is acceptable according to the Codex Allimentarius Commission (2010). DNA, with as low as 5 pg/μl, could be detected using q-PCR with primer of CYTBWB2-wb. The developed method was also used for DNA analysis extracted from meatball samples commercially available. Conclusion: q-PCR using CYTBWB2-wb primers targeting on mitochondrial cytochrome-b gene (forward: CGG TTC CCT CTT AGG CAT TT; Reverse: GGA TGA ACA GGC AGA TGA AGA) can be fruitfully used for the analysis of WBM in commercial meatball samples. [J Adv Vet Anim Res 2019; 6(3.000): 300-307]http://www.ejmanager.com/fulltextpdf.php?mno=17408CYTBWB2-wb primer; meatball; halal authentication; q-PCR; wild boar |
| spellingShingle | Ganea Qorry Aina Yuny Erwanto Motalib Hossain Mohd Rafie Johan Md. Eaqub Ali Abdul Rohman The employment of q-PCR using specific primer targeting on mitochondrial cytochrome-b gene for identification of wild boar meat in meatball samples Journal of Advanced Veterinary and Animal Research CYTBWB2-wb primer; meatball; halal authentication; q-PCR; wild boar |
| title | The employment of q-PCR using specific primer targeting on mitochondrial cytochrome-b gene for identification of wild boar meat in meatball samples |
| title_full | The employment of q-PCR using specific primer targeting on mitochondrial cytochrome-b gene for identification of wild boar meat in meatball samples |
| title_fullStr | The employment of q-PCR using specific primer targeting on mitochondrial cytochrome-b gene for identification of wild boar meat in meatball samples |
| title_full_unstemmed | The employment of q-PCR using specific primer targeting on mitochondrial cytochrome-b gene for identification of wild boar meat in meatball samples |
| title_short | The employment of q-PCR using specific primer targeting on mitochondrial cytochrome-b gene for identification of wild boar meat in meatball samples |
| title_sort | employment of q pcr using specific primer targeting on mitochondrial cytochrome b gene for identification of wild boar meat in meatball samples |
| topic | CYTBWB2-wb primer; meatball; halal authentication; q-PCR; wild boar |
| url | http://www.ejmanager.com/fulltextpdf.php?mno=17408 |
| work_keys_str_mv | AT ganeaqorryaina theemploymentofqpcrusingspecificprimertargetingonmitochondrialcytochromebgeneforidentificationofwildboarmeatinmeatballsamples AT yunyerwanto theemploymentofqpcrusingspecificprimertargetingonmitochondrialcytochromebgeneforidentificationofwildboarmeatinmeatballsamples AT motalibhossain theemploymentofqpcrusingspecificprimertargetingonmitochondrialcytochromebgeneforidentificationofwildboarmeatinmeatballsamples AT mohdrafiejohan theemploymentofqpcrusingspecificprimertargetingonmitochondrialcytochromebgeneforidentificationofwildboarmeatinmeatballsamples AT mdeaqubali theemploymentofqpcrusingspecificprimertargetingonmitochondrialcytochromebgeneforidentificationofwildboarmeatinmeatballsamples AT abdulrohman theemploymentofqpcrusingspecificprimertargetingonmitochondrialcytochromebgeneforidentificationofwildboarmeatinmeatballsamples AT ganeaqorryaina employmentofqpcrusingspecificprimertargetingonmitochondrialcytochromebgeneforidentificationofwildboarmeatinmeatballsamples AT yunyerwanto employmentofqpcrusingspecificprimertargetingonmitochondrialcytochromebgeneforidentificationofwildboarmeatinmeatballsamples AT motalibhossain employmentofqpcrusingspecificprimertargetingonmitochondrialcytochromebgeneforidentificationofwildboarmeatinmeatballsamples AT mohdrafiejohan employmentofqpcrusingspecificprimertargetingonmitochondrialcytochromebgeneforidentificationofwildboarmeatinmeatballsamples AT mdeaqubali employmentofqpcrusingspecificprimertargetingonmitochondrialcytochromebgeneforidentificationofwildboarmeatinmeatballsamples AT abdulrohman employmentofqpcrusingspecificprimertargetingonmitochondrialcytochromebgeneforidentificationofwildboarmeatinmeatballsamples |