Affiliations 

  • 1 Faculty of Applied Science and Technology, Universiti Tun Hussein Onn Malaysia, Hub Pendidikan Tinggi Pagoh, Muar 84600, Malaysia
  • 2 Halal Products Research Institute, Universiti Putra Malaysia, Putra Infoport, Serdang 43400, Malaysia
  • 3 Nanotechnology and Catalysis Research Centre, Institute for Advanced Studies, Universiti Malaya, Kuala Lumpur 50603, Malaysia
  • 4 Faculty of Food Science and Nutrition, Universiti Malaysia Sabah, Jalan UMS, Kota Kinabalu 88400, Malaysia
Molecules, 2022 Nov 22;27(23).
PMID: 36500215 DOI: 10.3390/molecules27238122

Abstract

Point-of-care diagnostic methods for animal species determination are critical for rapid, simple, and accurate enforcement of food labelling. PCR is the most common method for species identification. However, the requirement of using a thermal cycler created drawbacks for the PCR application, particularly in low-resource settings. Hence, in this study, a method for porcine DNA detection using recombinase polymerase amplification (RPA), coupled with nucleic acid lateral flow immunoassay (NALFIA), was developed. Porcine-specific primers targeting pig (Sus scrofa) cytochrome b gene fragments specifically amplify a 197 bp fragment of the mitochondrial gene as being visualized by 2% agarose gel and PCRD NALFIA. The reaction temperature and time were 39 °C and 20 min, respectively. Herein, the specificity of the primers to porcine was confirmed after being assayed against six animal species, namely cow, goat, chicken, duck, dog, and rabbit. The porcine-specific RPA assay shows a high limit of detection of 0.01 ng/µL pork DNA. Based on the preliminary performance data obtained from this study, the potential of this method as a rapid and sensitive tool for porcine DNA detection in meat-based products is foreseen.

* Title and MeSH Headings from MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.