Development of short amplicon-length PCR-RFLP assay for the detection of Macaca fascicularis meat under complex matrices / Nur Raifana binti Abdul Rashid

The macaque (Macaca fascicularis) monkeys are the third-largest primate population which are abundant in tropical forests. Despite being the potential carrier of Simian Immunodeficiency, Ebola and Corona viruses as well as religious and wildlife restrictions, macaques have been widely hunted and con...

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Main Author:	Abdul Rashid, Nur Raifana
Format:	Thesis
Published:	2015
Subjects:	Q Science (General) QD Chemistry QH301 Biology
Online Access:	http://studentsrepo.um.edu.my/5964/ http://studentsrepo.um.edu.my/5964/1/COMPLETE_THESIS_NUR_RAIFANA_HGA120006.pdf

id	um-stud-5964
recordtype	eprints
spelling	um-stud-59642015-10-07T08:19:47Z Development of short amplicon-length PCR-RFLP assay for the detection of Macaca fascicularis meat under complex matrices / Nur Raifana binti Abdul Rashid Abdul Rashid, Nur Raifana Q Science (General) QD Chemistry QH301 Biology The macaque (Macaca fascicularis) monkeys are the third-largest primate population which are abundant in tropical forests. Despite being the potential carrier of Simian Immunodeficiency, Ebola and Corona viruses as well as religious and wildlife restrictions, macaques have been widely hunted and consumed in many countries. However, in spite of being a potential adulterant of common meat, methods to detect monkey species in food are rarely documented. To fill up this research gap, here a monkey-specific polymerase chain reaction (PCR) assay targeting a short site (120bp) of mitochondrial d-loop gene was described since short-length targets are thermodynamically more stable than the longer ones under compromised states. The theoretical specificity of the primer pair was confirmed against 51 species, including 34 primates of which 13 species were from Macaque genera. The primers were fairly conserved for most of the Macaques but greatly polymorphic for other primates, demonstrating its universal signature for macaque detection. However, due to wildlife restriction, the practical specificity was tested only against 17 terrestrial and aquaticspecies and no cross-species amplification was detected under raw, processed and admixed states. The sensitivity of the assay was 0.0001ng DNA under pure states and 0.1% monkey meat in binary meat mixtures. Finally, the assay was validated by digesting the PCR products with AluI and CViKI-1 and distinctive restriction fingerprints for macaque identification were demonstrated both under raw meat 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 microfluidic lab-on-a-chip automated electrophoretic system separated the fragments with high resolution. Definitely the assay would be useful to regulatory bodies for food and feeds along with wildlife protection agencies as a reliable authentication technique for the unambiguous tracing of monkey meat under various matrices including the processed food. 2015 Thesis NonPeerReviewed application/pdf http://studentsrepo.um.edu.my/5964/1/COMPLETE_THESIS_NUR_RAIFANA_HGA120006.pdf Abdul Rashid, Nur Raifana (2015) Development of short amplicon-length PCR-RFLP assay for the detection of Macaca fascicularis meat under complex matrices / Nur Raifana binti Abdul Rashid. Masters thesis, University of Malaya. http://studentsrepo.um.edu.my/5964/
repository_type	Digital Repository
institution_category	Local University
institution	University Malaya
building	UM Students Repository
collection	Online Access
topic	Q Science (General) QD Chemistry QH301 Biology
spellingShingle	Q Science (General) QD Chemistry QH301 Biology Abdul Rashid, Nur Raifana Development of short amplicon-length PCR-RFLP assay for the detection of Macaca fascicularis meat under complex matrices / Nur Raifana binti Abdul Rashid
description	The macaque (Macaca fascicularis) monkeys are the third-largest primate population which are abundant in tropical forests. Despite being the potential carrier of Simian Immunodeficiency, Ebola and Corona viruses as well as religious and wildlife restrictions, macaques have been widely hunted and consumed in many countries. However, in spite of being a potential adulterant of common meat, methods to detect monkey species in food are rarely documented. To fill up this research gap, here a monkey-specific polymerase chain reaction (PCR) assay targeting a short site (120bp) of mitochondrial d-loop gene was described since short-length targets are thermodynamically more stable than the longer ones under compromised states. The theoretical specificity of the primer pair was confirmed against 51 species, including 34 primates of which 13 species were from Macaque genera. The primers were fairly conserved for most of the Macaques but greatly polymorphic for other primates, demonstrating its universal signature for macaque detection. However, due to wildlife restriction, the practical specificity was tested only against 17 terrestrial and aquaticspecies and no cross-species amplification was detected under raw, processed and admixed states. The sensitivity of the assay was 0.0001ng DNA under pure states and 0.1% monkey meat in binary meat mixtures. Finally, the assay was validated by digesting the PCR products with AluI and CViKI-1 and distinctive restriction fingerprints for macaque identification were demonstrated both under raw meat 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 microfluidic lab-on-a-chip automated electrophoretic system separated the fragments with high resolution. Definitely the assay would be useful to regulatory bodies for food and feeds along with wildlife protection agencies as a reliable authentication technique for the unambiguous tracing of monkey meat under various matrices including the processed food.
format	Thesis
author	Abdul Rashid, Nur Raifana
author_facet	Abdul Rashid, Nur Raifana
author_sort	Abdul Rashid, Nur Raifana
title	Development of short amplicon-length PCR-RFLP assay for the detection of Macaca fascicularis meat under complex matrices / Nur Raifana binti Abdul Rashid
title_short	Development of short amplicon-length PCR-RFLP assay for the detection of Macaca fascicularis meat under complex matrices / Nur Raifana binti Abdul Rashid
title_full	Development of short amplicon-length PCR-RFLP assay for the detection of Macaca fascicularis meat under complex matrices / Nur Raifana binti Abdul Rashid
title_fullStr	Development of short amplicon-length PCR-RFLP assay for the detection of Macaca fascicularis meat under complex matrices / Nur Raifana binti Abdul Rashid
title_full_unstemmed	Development of short amplicon-length PCR-RFLP assay for the detection of Macaca fascicularis meat under complex matrices / Nur Raifana binti Abdul Rashid
title_sort	development of short amplicon-length pcr-rflp assay for the detection of macaca fascicularis meat under complex matrices / nur raifana binti abdul rashid
publishDate	2015
url	http://studentsrepo.um.edu.my/5964/ http://studentsrepo.um.edu.my/5964/1/COMPLETE_THESIS_NUR_RAIFANA_HGA120006.pdf
first_indexed	2018-09-06T08:19:37Z
last_indexed	2018-09-06T08:19:37Z
_version_	1610845495939301376

Development of short amplicon-length PCR-RFLP assay for the detection of Macaca fascicularis meat under complex matrices / Nur Raifana binti Abdul Rashid

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