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Lab-on-a-Chip PCR-RFLP Assay for the Detection of Canine DNA in Burger Formulations

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Abstract

Canine species detection in foods is important in the perspectives of health, religions, and fare-trade food business. This study describes a very short-amplicon length Polymerase Chain Reaction (PCR)-Restriction Fragment Length Polymorphism (RFLP) assay with lab-on-a-chip detection platform for the authentication of canine DNA in processed foods. A 100-bp fragment of canine mitochondrial Cytochrome b (cytb) gene was selected and amplified using a pair of canine-specific primers. The amplified PCR products were validated by RFLP analysis using lab-on-a-chip microfluidic bioanalyzer kit. Both gel-image and electropherograms authenticated the canine-specific PCR products before (100 bp) and after restriction digestion (51, 30, and 19 bp). The assay successfully detected 0.0001-ng canine DNA under pure state and 0.01 % (w/w) canine meat spiked in chicken and beef burger formulations. Screening of eight commercial burgers across Malaysia did not reveal any canine adulteration. We believe the assay would find potential applications in food industries, Halal food regulatory bodies and animal right protection authorities across the globe.

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Acknowledgments

This research was jointly supported by University of Malaya research grant no. GC001A-14SBS to M.E. Ali and grant no: RU002-2014 to S.B.A. Hamid. M.M. Rahman is a recipient of Malaysian International Scholarship no. KPT.B.600-18/3 Vol2 133.

Conflict of Interest

Md. Eaqub Ali declares that he received funding support from the University of Malaya, supervised this work, and edited manuscript, and he has no conflict of interest to publish this paper. Md. Mahfujur Rahman declares that he performed this work under the supervision of Md. Eaqub Ali, and he has no conflict of interest to publish this paper. Sharifah Bee Abd Hamid declares that she paid salary to Md. Mahfujur Rahman from her research grant, and she has no conflict of interest to publish this paper. Subha Bhassu declares that she provided some reagents to Md. Mahfujur Rahman, and she has no conflict of interest to publish this paper. Shuhaimi Mustafa declares that he ensured lab facilities for this work, and he has no conflict of interest to publish this paper. Md. Al Amin declares that he helped Md. Mahfujur Rahman in sample collection, and he has no conflict of interest to publish this paper. Md. Abdur Razzak declares that he helped Md. Mahfujur Rahman in experiments, and he has no conflict of interest to publish this paper.

Compliance with Ethics Requirements

Ethical clearance of ref. no: NANOCAT /25/04/3013/ MMR (R) was obtained from the Institutional Animal Care and Use Committee, University of Malaya (UM IACUC), and all experiments were conducted and animal meats were handled following the national and institutional guideline.

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Authors and Affiliations

  1. Nanotechnology and Catalysis Research Centre, University of Malaya, Kuala Lumpur, 50603, Malaysia

    Md. Mahfujur Rahman, Md. Eaqub Ali, Sharifah Bee Abd Hamid, Md. Al Amin & Md. Abdur Razzak

  2. Institute of Biological Sciences, University of Malaya, Kuala Lumpur, 50603, Malaysia

    Subha Bhassu

  3. CEBAR Laboratory, IPS Building, University of Malaya, Kuala Lumpur, 50603, Malaysia

    Md. Eaqub Ali & Subha Bhassu

  4. Institute of Halal Products Research, University of Putra Malaysia, Serdang, 43400, Malaysia

    Shuhaimi Mustafa

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  1. Md. Mahfujur Rahman
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Correspondence to Md. Eaqub Ali.

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Rahman, M.M., Ali, M.E., Hamid, S.B.A. et al. Lab-on-a-Chip PCR-RFLP Assay for the Detection of Canine DNA in Burger Formulations. Food Anal. Methods 8, 1598–1606 (2015). https://doi.org/10.1007/s12161-015-0090-1

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