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Double gene targeting multiplex PCR-RFLP detects Crocodylus porosus in chicken meatball and traditional medicine

Crocodiles have been hunted and consumed for centuries for skins, nutrients, and medicines. These indomitable trends have overpowered restrictions from wildlife and conservation agencies, continuing the illegal trades of crocodiles across the world. This paper described the development of a very sta...

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Main Authors: Ahmad Nizar, Nina Naquiah, Sultana, Sharmin, Hossain, M.A. Motalib, Johan, Mohd Rafie, Ali, Md. Eaqub
Format: Article
Published: Taylor & Francis 2018
Subjects:
Q Science (General)
R Medicine
Online Access:http://eprints.um.edu.my/21121/
https://doi.org/10.1080/10942912.2018.1508164
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spelling my.um.eprints.211212019-05-03T08:20:40Z http://eprints.um.edu.my/21121/ Double gene targeting multiplex PCR-RFLP detects Crocodylus porosus in chicken meatball and traditional medicine Ahmad Nizar, Nina Naquiah Sultana, Sharmin Hossain, M.A. Motalib Johan, Mohd Rafie Ali, Md. Eaqub Q Science (General) R Medicine Crocodiles have been hunted and consumed for centuries for skins, nutrients, and medicines. These indomitable trends have overpowered restrictions from wildlife and conservation agencies, continuing the illegal trades of crocodiles across the world. This paper described the development of a very stable, fast, and secured polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay for the confirmed detection of Crocodylus porosus under any matrices and decomposing treatments. Two very short-sites (77 and 127-bp) of atp6 and cytb genes of C. porosus were controlled digested with AciI enzyme; producing distinctive RFLP patterns (83, 54, 44 & 23 bp). The enzyme digested assay was stable following extreme boiling, autoclaving, and microwaving treatments that break down DNA. The sensitivity was tested and validated in model meatballs and it was suitable for detecting 0.01% crocodile meatball matrices. The optimized RFLP assay was used to screen 3 commercial meatballs and 21 traditional medicines (TM). While no crocodile DNA was found in commercial chicken meatballs, 4/21 TM products were found correctly labelled to contain C. porosus DNA. The novel assay demonstrated sufficient merit to be used by regulatory agencies for any forensic and/or archaeological identification of C. porosus even under the state of decomposition. Taylor & Francis 2018 Article PeerReviewed Ahmad Nizar, Nina Naquiah and Sultana, Sharmin and Hossain, M.A. Motalib and Johan, Mohd Rafie and Ali, Md. Eaqub (2018) Double gene targeting multiplex PCR-RFLP detects Crocodylus porosus in chicken meatball and traditional medicine. International Journal of Food Properties, 21 (1). pp. 2037-2051. ISSN 1094-2912 https://doi.org/10.1080/10942912.2018.1508164 doi:10.1080/10942912.2018.1508164
institution Universiti Malaya
building UM Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Malaya
content_source UM Research Repository
url_provider http://eprints.um.edu.my/
topic Q Science (General)
R Medicine
spellingShingle Q Science (General)
R Medicine
Ahmad Nizar, Nina Naquiah
Sultana, Sharmin
Hossain, M.A. Motalib
Johan, Mohd Rafie
Ali, Md. Eaqub
Double gene targeting multiplex PCR-RFLP detects Crocodylus porosus in chicken meatball and traditional medicine
description Crocodiles have been hunted and consumed for centuries for skins, nutrients, and medicines. These indomitable trends have overpowered restrictions from wildlife and conservation agencies, continuing the illegal trades of crocodiles across the world. This paper described the development of a very stable, fast, and secured polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay for the confirmed detection of Crocodylus porosus under any matrices and decomposing treatments. Two very short-sites (77 and 127-bp) of atp6 and cytb genes of C. porosus were controlled digested with AciI enzyme; producing distinctive RFLP patterns (83, 54, 44 & 23 bp). The enzyme digested assay was stable following extreme boiling, autoclaving, and microwaving treatments that break down DNA. The sensitivity was tested and validated in model meatballs and it was suitable for detecting 0.01% crocodile meatball matrices. The optimized RFLP assay was used to screen 3 commercial meatballs and 21 traditional medicines (TM). While no crocodile DNA was found in commercial chicken meatballs, 4/21 TM products were found correctly labelled to contain C. porosus DNA. The novel assay demonstrated sufficient merit to be used by regulatory agencies for any forensic and/or archaeological identification of C. porosus even under the state of decomposition.
format Article
author Ahmad Nizar, Nina Naquiah
Sultana, Sharmin
Hossain, M.A. Motalib
Johan, Mohd Rafie
Ali, Md. Eaqub
author_facet Ahmad Nizar, Nina Naquiah
Sultana, Sharmin
Hossain, M.A. Motalib
Johan, Mohd Rafie
Ali, Md. Eaqub
author_sort Ahmad Nizar, Nina Naquiah
title Double gene targeting multiplex PCR-RFLP detects Crocodylus porosus in chicken meatball and traditional medicine
title_short Double gene targeting multiplex PCR-RFLP detects Crocodylus porosus in chicken meatball and traditional medicine
title_full Double gene targeting multiplex PCR-RFLP detects Crocodylus porosus in chicken meatball and traditional medicine
title_fullStr Double gene targeting multiplex PCR-RFLP detects Crocodylus porosus in chicken meatball and traditional medicine
title_full_unstemmed Double gene targeting multiplex PCR-RFLP detects Crocodylus porosus in chicken meatball and traditional medicine
title_sort double gene targeting multiplex pcr-rflp detects crocodylus porosus in chicken meatball and traditional medicine
publisher Taylor & Francis
publishDate 2018
url http://eprints.um.edu.my/21121/
https://doi.org/10.1080/10942912.2018.1508164
_version_ 1643691472007987200
score 13.211869

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