This review describes all the available DNA methods, which can detect pork in foods, as a basis for Halal verification. The review describes the following assays: Singleplex and Multiplex PCR, PCR-RAPD and PCR-RFLP, Real Time-PCR, DNA-Barcoding, Loop-mediated isothermal amplification (LAMP), Biosensor - optic and electrochemical, and CRISPR/Cas12. It discusses the detail probes, primers etc as well as the advantages and disadvantages of each method.
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In the Islamic and Jewish religions certain animals and seafood are forbidden. Pork is forbidden to eat for both religions, and in Judaism, only animals which have cloven hooves and chew the cud can be consumed. This review looks at the different methods and techniques to determine the species of meat, or meat based ingredients and derived fats in halal/kosher foods. It does not address the other aspect of halal/kosher foods, namely the method of slaughter of the animal.
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In this paper, a new approach to rapid pork detection was developed using an amplification-free and mix-to-read CRISPR-Cas12-based nucleic acid analytical strategy. An optimized guide RNA (gRNA) targeting the pork cytochrome b (Cyt b) gene was designed, which allowed specific identification of the target Cyt b gene in pork components. Activation of Cas12 protein to cleave single-stranded DNA probes with terminally labelled fluorophore and quencher groups then allowed confirmation of the presence of pork Cytb by reading the fluorescence signal. The assay allowed specific detection of pork in beef, mutton, and chicken products, The reliability of the method was tested on processed halal meat products - beef luncheon meat and spiced beef, as well as non-halal foods - sausage and dried pork slices.
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The use of pork and porcine ingredients is banned in halal and kosher foods. This review by Indonesian researchers examines the various methods from DNA analysis, FTIR spectroscopic analysis, chromatography to electronic nose, that have been used to detect porcine DNA, pork, pork gelatine, and lard in meat products.
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