TaqMan probe based multiplex quantitative PCR assay for determination of bovine, porcine and fish DNA in gelatin admixture, food products and dietary supplements

Sharmin Sultana; M A Motalib Hossain; Azrina Azlan; Mohd Rafie Johan; Zaira Zaman Chowdhury; Md Eaqub Ali

doi:10.1016/j.foodchem.2020.126756

TaqMan probe based multiplex quantitative PCR assay for determination of bovine, porcine and fish DNA in gelatin admixture, food products and dietary supplements

Food Chem. 2020 Apr 7:325:126756. doi: 10.1016/j.foodchem.2020.126756. Online ahead of print.

Authors

Sharmin Sultana¹, M A Motalib Hossain¹, Azrina Azlan², Mohd Rafie Johan¹, Zaira Zaman Chowdhury¹, Md Eaqub Ali³

Affiliations

¹ Nanotechnology and Catalysis Research Centre, Institute for Advanced Studies, University of Malaya, 50603 Kuala Lumpur, Malaysia.
² Department of Nutrition & Dietetics, Faculty of Medicine & Health Sciences, University Putra Malaysia, Serdang, 43400 Selangor, Malaysia.
³ Nanotechnology and Catalysis Research Centre, Institute for Advanced Studies, University of Malaya, 50603 Kuala Lumpur, Malaysia; Centre for Research in Biotechnology for Agriculture, University of Malaya, 50603 Kuala Lumpur, Malaysia. Electronic address: eaqubali@um.edu.my.

PMID: 32413685
DOI: 10.1016/j.foodchem.2020.126756

Abstract

Detection of animal materials in gelatin-based products is required to address religious and cultural concerns, because porcine and bovine gelatins are prohibited in Halal, Kosher and Hindus consumer goods. In this paper, multiplex quantitative polymerase chain reaction (qPCR) assay using TaqMan probe was developed to discriminate bovine, porcine and fish gelatin species in a single assay platform. The assay was specific to cattle, pigs and fish, having been tested against 14 non-target species. The limit of detection, under gelatin admixed conditions, was 0.005 ng/µL. Finally, a pilot survey was undertaken testing 35 Halal branded processed food and dietary items. Out of 35 samples, only two were found to be positive for porcine species. The authenticity of these two qPCR products was confirmed by DNA sequencing analysis, which showed 99-100% similarity with Sus scrofa (Wild boar) species.

Keywords: Internal amplification control; Limit of quantification; Multiplex quantitative PCR; PCR efficiency; TaqMan Probes.