title: A suitable method for the detection of a potential fraud of bringing macaque monkey meat into the food chain.
abstract: Being the third-largest primate population has not made macaque (Macaca fascicularis sp.) monkeys less exposed to threats and dangers. Despite wildlife protection, they have been widely hunted and consumed in several countries because of their purported nutritional values. In addition to trading as pure bush meats in several places, monkey meat has been sold in meatball and soup products in Indonesia. Thus the possibility of macaque meat trafficking under the label of common meats is quite high. This paper reports the development of a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay with the shortest amplicon length for the confirmed detection of monkey meat under compromised states which are known to degrade DNA. We amplified a 120-bp region of d-loop gene using a pair of macaque-specific primers and confirmed their specificity for the target species through cross-challenging against 17 different species using a 141-bp site of an 18 S rRNA gene as an endogenous control for eukaryotes. This eliminated the possibilities of any false-negative detection with complex matrices or degraded specimens. The detection limit was 0.00001 ng DNA in a pure state and 0.1% of meat in mixed matrices and commercial meatball products. RFLP analysis further authenticated the originality of the PCR product and distinctive restriction patterns were found upon AluI and CViKI-1 digestion. A micro-fluidic lab-on-a-chip automated electrophoretic system separated the fragments with high resolution. The assay was validated for screening commercial meatball products with sufficient internal control.
authors.affiliation: a Nanotechnology and Catalysis Research Centre (NanoCat) , University of Malaya , Kuala Lumpur , Malaysia.
authors.forename: Nur Raifana Abdul
authors.initials: NR
authors.lastname: Rashid
authors.forename: Md Eaqub
authors.initials: ME
authors.lastname: Ali
authors.forename: Sharifah Bee Abd
authors.initials: SB
authors.lastname: Hamid
authors.forename: Md Mahfujur
authors.initials: MM
authors.lastname: Rahman
authors.forename: Md Abdur
authors.initials: MA
authors.lastname: Razzak
authors.lastname: Asing
authors.forename: Md Al
authors.initials: MA
authors.lastname: Amin
mesh_terms: D000818:Animals; D004247:DNA / Q000235:genetics*; D005504:Food Analysis / Q000379:methods*; D005506:Food Contamination; D005607:Fraud; D008251:Macaca; D008460:Meat / Q000032:analysis*; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D012150:Polymorphism, Restriction Fragment Length; D012680:Sensitivity and Specificity; D013045:Species Specificity
chemical_list: D004247:DNA
keywords: compromised states; endogenous control; macaque monkey; meat trafficking; reproducibility; restriction patterns