Detection of Dog (Canis lupus familiaris) and Pig (Sus scrofa) Meat Components for Halal Authentication using Duplex Loop-Mediated Isothermal Amplification (Du-LAMP)

HistoryThis study aimed to develop and validate a duplex loop-mediated isothermal amplification (Du-LAMP) assay for the early, simultaneous detection of dog (Canis lupus familiaris) and pig (Sus scrofa) DNA in food products to support Halal authentication efforts.Species-specific primers targeting mitochondrial DNA were designed for both species.The assay was tested on raw and cooked meat samples under laboratory conditions.Specificity, sensitivity, and timeto-result were evaluated and compared against conventional PCR.Negative controls included Diethyl pyrocarbonate (DEPC)-treated water and non-target species.The Du-LAMP assay specifically detected both dog and pig DNA, with no cross-reactivity.The limit of detection was 200 femtograms, approximately 50 times more sensitive than conventional PCR.Amplification was completed within 30min at 68°C, with visualization by agarose gel electrophoresis.In contrast, PCR required ~90min and thermal cycling.This study reports the first duplex LAMP assay capable of simultaneously detecting dog and pig DNA in a single reaction.The method is rapid, sensitive, and potentially adaptable for field use in Halal compliance screening.Further validation on commercial food products is recommended to support real-world applications.