TaqMan probe real-time polymerase chain reaction assay for the quantification of canine DNA in chicken nugget. Issue 1 (2nd January 2016)

• Record Type: Journal Article
• Authors: Rahman, Md. Mahfujur; Hamid, Sharifah Bee Abd; Basirun, Wan Jefrey; Bhassu, Subha; Rashid, Nur Raifana Abdul; Mustafa, Shuhaimi; Mohd Desa, Mohd Nasir; Ali, Md. Eaqub
• Is Part Of: Food additives & contaminants. Volume 33:Issue 1(2016)
• Page Start: 10
• Page End: 18
• Publication Date: 2016-01-02
• Subjects: qPCR assay -- canine species -- nugget formulation -- halal meat -- food adulteration; Food additives -- Periodicals; Food contamination -- Periodicals; 664.06
• DOI: 10.1080/19440049.2015.1104558
• Languages: English
• ISSNs: 1944-0049

Abstract

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.