Rapid detection of porcine DNA in meatball using recombinase polymerase amplification couple with lateral flow immunoassay for halal authentication

Mohd Hazim Mohd Yusop and Mohd Fadzelly Abu Bakar and Kamarul Rahim Kamarudin 1, and Nur Fadhilah Khairil Mokhtar and Mohd Abd Motalib Hossain and Mohd Rafie Johan and Nor Qhairul Izzreen Mohd Noor (2022) Rapid detection of porcine DNA in meatball using recombinase polymerase amplification couple with lateral flow immunoassay for halal authentication. Molecules, 27 (8122). pp. 1-9.

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URL: https://www.mdpi.com/1420-3049/27/23/8122

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.

Item Type:	Article
Keyword:	Porcine DNA, RPA, NALFIA, Meatball, Halal authentication
Subjects:	T Technology > TX Home economics > TX1-1110 Home economics > TX341-641 Nutrition. Foods and food supply
Department:	SCHOOL > School of Food Science and Nutrition
Depositing User:	DG MASNIAH AHMAD -
Date Deposited:	26 Sep 2023 10:14
Last Modified:	26 Sep 2023 10:14
URI:	https://eprints.ums.edu.my/id/eprint/37405

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