title: TaqMan probe real-time polymerase chain reaction assay for the quantification of canine DNA in chicken nugget.
abstract: This paper describes a short-amplicon-based TaqMan probe quantitative real-time PCR (qPCR) assay for the quantitative detection of canine meat in chicken nuggets, which are very popular across the world, including Malaysia. The assay targeted a 100-bp fragment of canine cytb gene using a canine-specific primer and TaqMan probe. Specificity against 10 different animals and plants species demonstrated threshold cycles (Ct) of 16.13 ± 0.12 to 16.25 ± 0.23 for canine DNA and negative results for the others in a 40-cycle reaction. The assay was tested for the quantification of up to 0.01% canine meat in deliberately spiked chicken nuggets with 99.7% PCR efficiency and 0.995 correlation coefficient. The analysis of the actual and qPCR predicted values showed a high recovery rate (from 87% ± 28% to 112% ± 19%) with a linear regression close to unity (R(2) = 0.999). Finally, samples of three halal-branded commercial chicken nuggets collected from different Malaysian outlets were screened for canine meat, but no contamination was demonstrated.
authors.affiliation: a Nanotechnology and Catalysis Research Centre , University of Malayaob , Kuala Lumpur , Malaysia.
authors.forename: Md Mahfujur
authors.initials: MM
authors.lastname: Rahman
authors.affiliation: a Nanotechnology and Catalysis Research Centre , University of Malayaob , Kuala Lumpur , Malaysia.
authors.forename: Sharifah Bee Abd
authors.initials: SB
authors.lastname: Hamid
authors.affiliation: a Nanotechnology and Catalysis Research Centre , University of Malayaob , Kuala Lumpur , Malaysia.|b Department of Chemistry , University of Malaya , Kuala Lumpur , Malaysia.
authors.forename: Wan Jefrey
authors.initials: WJ
authors.lastname: Basirun
authors.affiliation: c Institute of Biological Sciences & CEBAR Laboratory , University of Malaya , Kuala Lumpur , Malaysia.
authors.forename: Subha
authors.initials: S
authors.lastname: Bhassu
authors.affiliation: a Nanotechnology and Catalysis Research Centre , University of Malayaob , Kuala Lumpur , Malaysia.
authors.forename: Nur Raifana Abdul
authors.initials: NR
authors.lastname: Rashid
authors.affiliation: d Institute of Halal Products Research , University of Putra Malaysia , Serdang , Malaysia.
authors.forename: Shuhaimi
authors.initials: S
authors.lastname: Mustafa
authors.affiliation: d Institute of Halal Products Research , University of Putra Malaysia , Serdang , Malaysia.
authors.forename: Mohd Nasir
authors.initials: MN
authors.lastname: Mohd Desa
authors.affiliation: a Nanotechnology and Catalysis Research Centre , University of Malayaob , Kuala Lumpur , Malaysia.
authors.forename: Md Eaqub
authors.initials: ME
authors.lastname: Ali
mesh_terms: D000818:Animals; D002645:Chickens; D004247:DNA / Q000032:analysis* / Q000235:genetics; D004285:Dogs / Q000235:genetics*; D005506:Food Contamination / Q000032:analysis*; D008460:Meat / Q000032:analysis*; D015345:Oligonucleotide Probes / Q000032:analysis / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*
chemical_list: D015345:Oligonucleotide Probes; D004247:DNA
keywords: canine species; food adulteration; halal meat; nugget formulation; qPCR assay