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Rapid porcine detection in gelatin-based highly processed products using loop mediated isothermal amplification

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Abstract

Low DNA concentration recovered from highly processed products such as gelatin and gelatin-based products renders difficulty in detecting porcine contamination using conventional PCR techniques. We documented here a porcine-specific loop-mediated isothermal amplification (LAMP) to identify porcine traces in gelatin products. The porcine-specific primers were designed according to mitochondrial DNA of Cytochrome b gene sequence. Here we used two different reaction mixtures for LAMP assay (GENIE and MYRM) against the same DNA samples extracted from gelatin products and porcine-specific primers to detect the presence of porcine DNA. The porcine-specific primers were shown to be specific only to Sus scrofa against 14 DNA of other meat species. The analytical sensitivity of the LAMP assay for porcine DNA detection is 1 pg/µL using both GENIE (within 30 m) and MYRM (within 60 m) reaction mixtures. Analysis against 32 samples of gelatin products showed that five samples were found to contain porcine DNA; two samples out of six gelatin powder samples and three gelatin capsule samples out of nine. Out of these five positive samples, three were not labeled containing porcine gelatin. Overall, LAMP assay in this study showed an excellent specificity, sensitivity and rapidity in detection of porcine DNA in gelatin products.

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Acknowledgements

The authors would like to acknowledge the financial support provided by Universiti Putra Malaysia and Konsortium Institut Halal IPT Malaysia. Nor Asmara Tasrip wishes to acknowledge the financial support from MyBrain15 under the Ministry of Higher Education, Malaysia for her PhD fellowship.

Funding

This study was funded by grants of Universiti Putra Malaysia (GP-IPS/2018/9625800 and GP-B/2018/9658200) and Konsortium Institut Halal IPT Malaysia, Ministry of Higher Education, Malaysia (6300911).

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Authors and Affiliations

  1. Halal Products Research Institute, Universiti Putra Malaysia, 43400, Serdang, Selangor, Malaysia

    Nor Asmara Tasrip, Mohd Nasir Mohd Desa, Nur Fadhilah Khairil Mokhtar & Nurhayatie Sajali

  2. Ministry of Higher Education, Konsortium Institut Halal IPT Malaysia, 62200, Putrajaya, Malaysia

    Mohd Nasir Mohd Desa, Nur Fadhilah Khairil Mokhtar & Amalia Mohd Hashim

  3. Department of Biomedical Science, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, 43400, Serdang, Selangor, Malaysia

    Mohd Nasir Mohd Desa & Cheah Yoke Kqueen

  4. School of Engineering and Technology, University College of Technology Sarawak, 96000, Sibu, Sarawak, Malaysia

    Nurhayatie Sajali

  5. Department of Microbiology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400, Serdang, Selangor, Malaysia

    Amalia Mohd Hashim

  6. Modesto City Schools, 426 Locust St., Modesto, CA, 95351-2699, USA

    Md. Eaqub Ali

  7. Nanotechnology and Catalysis Research Center, University of Malaya, 50603, Kuala Lumpur, Malaysia

    Md. Eaqub Ali

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  1. Nor Asmara Tasrip
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Correspondence to Mohd Nasir Mohd Desa.

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Nor Asmara Tasrip declares that she has no conflict of interest. Mohd Nasir Mohd Desa declares that he has no conflict of interest. Nur Fadhilah Khairil Mokhtar declares that she has no conflict of interest. Nurhayatie Sajali declares that she has no conflict of interest. Amalia Mohd Hashim declares that she has no conflict of interest. Md. Eaqub Ali declares that he has no conflict of interest. Cheah Yoke Kqueen declares that he has no conflict of interest.

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Tasrip, N.A., Mohd Desa, M.N., Khairil Mokhtar, N.F. et al. Rapid porcine detection in gelatin-based highly processed products using loop mediated isothermal amplification. J Food Sci Technol 58, 4504–4513 (2021). https://doi.org/10.1007/s13197-020-04932-2

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