Development and validation of short-amplicon length PCR assay for macaques meat detection under complex matrices
2017
Ali, Md Eaqub | Raifana Abdul Rashid, Nur | Bee Abd Hamid, Sharifah | Hossain, S. M Azad | Asing, Asing | Hossain, M. A Motalib | Zaidul, I. S. M.
The macaque (Macaca fascicularis) monkeys are the third largest primate population in tropical forests. Being the potential carrier of Simian Immunodeficiency and Ebola and Corona viruses, as well as being a religiously-protected and wildlife-protected species could not save the macaques from being over-hunted and consumed. Despite having needs, methods to detect monkey species in foods are rarely documented. To fill this gap, this article describes a monkey-specific polymerase chain reaction assay targeting a short-site (120 bp) of mitochondrial d-loop gene because the short-length targets are thermodynamically more stable than the longer ones under degrading states. Specificity was tested against 17 terrestrial and aquatic meat and fish species and no cross-species amplification was detected under raw, processed, and admixed states. The sensitivity of the assay was 0.0001 ng DNA under pure states and 0.1% monkey meats in binary meat mixtures. Finally, the assay was validated by digesting the polymerase chain reaction products with Alu I and CViKI- 1 and distinctive restriction fingerprints for macaque species were demonstrated.
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