Duplex real-time PCR assay using SYBR Green to detect and quantify Malayan box turtle (Cuora amboinensis) materials in meatballs, burgers, frankfurters and traditional Chinese herbal jelly powder

The Malayan box turtle (Cuora amboinensis) (MBT) is a vulnerable and protected species widely used in exotic foods and traditional medicines. Currently available polymerase chain reaction (PCR) assays to identify MBT lack automation and involve long targets which break down in processed or denatured...

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Main Authors:	Asing, Md. Eaqub Ali, Ali, Md Eaqub, Hossain, Md. Motalib, Ahamad, Mohammad Nasir Uddin, Hossain, S.M. Azad, Ahmad Nizar, Nina Naquiah, Sarker, Md. Zaidul Islam
Format:	Article
Language:	English English English
Published:	Taylor & Francis 2016
Subjects:	Q Science (General)
Online Access:	http://irep.iium.edu.my/55692/ http://irep.iium.edu.my/55692/ http://irep.iium.edu.my/55692/ http://irep.iium.edu.my/55692/1/55692_Duplex%20real-time%20PCR%20assay%20using%20SYBR.pdf http://irep.iium.edu.my/55692/2/55692_Duplex%20real-time%20PCR%20assay%20using%20SYBR_SCOPUS.pdf http://irep.iium.edu.my/55692/3/55692_Duplex%20real-time%20PCR%20assay%20using%20SYBR_WOS.pdf

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spelling	iium-556922017-10-12T01:51:26Z http://irep.iium.edu.my/55692/ Duplex real-time PCR assay using SYBR Green to detect and quantify Malayan box turtle (Cuora amboinensis) materials in meatballs, burgers, frankfurters and traditional Chinese herbal jelly powder Asing, Md. Eaqub Ali Ali, Md Eaqub Hossain, Md. Motalib Ahamad, Mohammad Nasir Uddin Hossain, S.M. Azad Ahmad Nizar, Nina Naquiah Sarker, Md. Zaidul Islam Q Science (General) The Malayan box turtle (Cuora amboinensis) (MBT) is a vulnerable and protected species widely used in exotic foods and traditional medicines. Currently available polymerase chain reaction (PCR) assays to identify MBT lack automation and involve long targets which break down in processed or denatured tissue. This SYBR Green duplex real-time PCR assay has addressed this research gap for the first time through the combination of 120- and 141-bp targets from MBT and eukaryotes for the quantitative detection of MBT DNA in food chain and herbal medicinal preparations. This authentication ensures better security through automation, internal control and short targets that were stable under the processing treatments of foods and medicines. A melting curve clearly demonstrated two peaks at 74.63 ± 0.22 and 78.40 ± 0.31°C for the MBT and eukaryotic products, respectively, under pure, admixed and commercial food matrices. Analysis of 125 reference samples reflected a target recovery of 93.25–153.00%, PCR efficiency of 99–100% and limit of detection of 0.001% under various matrices. The quantification limits were 0.00001, 0.00170 ± 0.00012, 0.00228 ± 0.00029, 0.00198 ± 0.00036 and 0.00191 ± 0.00043 ng DNA for the pure meat, binary mixtures, meatball, burger and frankfurter products, respectively. The assay was used to screen 100 commercial samples of traditional Chinese herbal jelly powder from eight different brands; 22% of them were found to be MBT-positive (5.37 ± 0.50–7.00 ± 0.34% w/w), which was reflected through the Ct values (26.37 ± 0.32–28.90 ± 0.42) and melting curves (74.63– 78.65 ± 0.22°C) of the amplified MBT target (120 bp), confirming the speculation that MBT materials are widely used in Chinese herbal desserts, exotic dishes consumed with the hope of prolonging life and youth. Taylor & Francis 2016-09-02 Article PeerReviewed application/pdf en http://irep.iium.edu.my/55692/1/55692_Duplex%20real-time%20PCR%20assay%20using%20SYBR.pdf application/pdf en http://irep.iium.edu.my/55692/2/55692_Duplex%20real-time%20PCR%20assay%20using%20SYBR_SCOPUS.pdf application/pdf en http://irep.iium.edu.my/55692/3/55692_Duplex%20real-time%20PCR%20assay%20using%20SYBR_WOS.pdf Asing, Md. Eaqub Ali and Ali, Md Eaqub and Hossain, Md. Motalib and Ahamad, Mohammad Nasir Uddin and Hossain, S.M. Azad and Ahmad Nizar, Nina Naquiah and Sarker, Md. Zaidul Islam (2016) Duplex real-time PCR assay using SYBR Green to detect and quantify Malayan box turtle (Cuora amboinensis) materials in meatballs, burgers, frankfurters and traditional Chinese herbal jelly powder. Food Additives and Contaminants - Part A Chemistry, Analysis, Control, Exposure and Risk Assessment, 33 (11). pp. 1643-1659. ISSN 1944-0049 E-ISSN 1944-0057 http://download.xuebalib.com/xuebalib.com.10952.pdf 10.1080/19440049.2016.1236403
repository_type	Digital Repository
institution_category	Local University
institution	International Islamic University Malaysia
building	IIUM Repository
collection	Online Access
language	English English English
topic	Q Science (General)
spellingShingle	Q Science (General) Asing, Md. Eaqub Ali Ali, Md Eaqub Hossain, Md. Motalib Ahamad, Mohammad Nasir Uddin Hossain, S.M. Azad Ahmad Nizar, Nina Naquiah Sarker, Md. Zaidul Islam Duplex real-time PCR assay using SYBR Green to detect and quantify Malayan box turtle (Cuora amboinensis) materials in meatballs, burgers, frankfurters and traditional Chinese herbal jelly powder
description	The Malayan box turtle (Cuora amboinensis) (MBT) is a vulnerable and protected species widely used in exotic foods and traditional medicines. Currently available polymerase chain reaction (PCR) assays to identify MBT lack automation and involve long targets which break down in processed or denatured tissue. This SYBR Green duplex real-time PCR assay has addressed this research gap for the first time through the combination of 120- and 141-bp targets from MBT and eukaryotes for the quantitative detection of MBT DNA in food chain and herbal medicinal preparations. This authentication ensures better security through automation, internal control and short targets that were stable under the processing treatments of foods and medicines. A melting curve clearly demonstrated two peaks at 74.63 ± 0.22 and 78.40 ± 0.31°C for the MBT and eukaryotic products, respectively, under pure, admixed and commercial food matrices. Analysis of 125 reference samples reflected a target recovery of 93.25–153.00%, PCR efficiency of 99–100% and limit of detection of 0.001% under various matrices. The quantification limits were 0.00001, 0.00170 ± 0.00012, 0.00228 ± 0.00029, 0.00198 ± 0.00036 and 0.00191 ± 0.00043 ng DNA for the pure meat, binary mixtures, meatball, burger and frankfurter products, respectively. The assay was used to screen 100 commercial samples of traditional Chinese herbal jelly powder from eight different brands; 22% of them were found to be MBT-positive (5.37 ± 0.50–7.00 ± 0.34% w/w), which was reflected through the Ct values (26.37 ± 0.32–28.90 ± 0.42) and melting curves (74.63– 78.65 ± 0.22°C) of the amplified MBT target (120 bp), confirming the speculation that MBT materials are widely used in Chinese herbal desserts, exotic dishes consumed with the hope of prolonging life and youth.
format	Article
author	Asing, Md. Eaqub Ali Ali, Md Eaqub Hossain, Md. Motalib Ahamad, Mohammad Nasir Uddin Hossain, S.M. Azad Ahmad Nizar, Nina Naquiah Sarker, Md. Zaidul Islam
author_facet	Asing, Md. Eaqub Ali Ali, Md Eaqub Hossain, Md. Motalib Ahamad, Mohammad Nasir Uddin Hossain, S.M. Azad Ahmad Nizar, Nina Naquiah Sarker, Md. Zaidul Islam
author_sort	Asing, Md. Eaqub Ali
title	Duplex real-time PCR assay using SYBR Green to detect and quantify Malayan box turtle (Cuora amboinensis) materials in meatballs, burgers, frankfurters and traditional Chinese herbal jelly powder
title_short	Duplex real-time PCR assay using SYBR Green to detect and quantify Malayan box turtle (Cuora amboinensis) materials in meatballs, burgers, frankfurters and traditional Chinese herbal jelly powder
title_full	Duplex real-time PCR assay using SYBR Green to detect and quantify Malayan box turtle (Cuora amboinensis) materials in meatballs, burgers, frankfurters and traditional Chinese herbal jelly powder
title_fullStr	Duplex real-time PCR assay using SYBR Green to detect and quantify Malayan box turtle (Cuora amboinensis) materials in meatballs, burgers, frankfurters and traditional Chinese herbal jelly powder
title_full_unstemmed	Duplex real-time PCR assay using SYBR Green to detect and quantify Malayan box turtle (Cuora amboinensis) materials in meatballs, burgers, frankfurters and traditional Chinese herbal jelly powder
title_sort	duplex real-time pcr assay using sybr green to detect and quantify malayan box turtle (cuora amboinensis) materials in meatballs, burgers, frankfurters and traditional chinese herbal jelly powder
publisher	Taylor & Francis
publishDate	2016
url	http://irep.iium.edu.my/55692/ http://irep.iium.edu.my/55692/ http://irep.iium.edu.my/55692/ http://irep.iium.edu.my/55692/1/55692_Duplex%20real-time%20PCR%20assay%20using%20SYBR.pdf http://irep.iium.edu.my/55692/2/55692_Duplex%20real-time%20PCR%20assay%20using%20SYBR_SCOPUS.pdf http://irep.iium.edu.my/55692/3/55692_Duplex%20real-time%20PCR%20assay%20using%20SYBR_WOS.pdf
first_indexed	2023-09-18T21:18:39Z
last_indexed	2023-09-18T21:18:39Z
_version_	1777411732676804608

Duplex real-time PCR assay using SYBR Green to detect and quantify Malayan box turtle (Cuora amboinensis) materials in meatballs, burgers, frankfurters and traditional Chinese herbal jelly powder

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