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A higher sensitivity and efficiency of common primer multiplex PCR assay in identification of meat origin using NADH dehydrogenase subunit 4 gene

A Common Primer Multiplex PCR (CP-M-PCR) was developed to detect meat origin of four groups of animal (pig, ruminant, avian and rabbit). This method demonstrated higher sensitivity and efficiency than the conventional multiplex PCR. In this approach, a common forward primer was designed in the 5′ en...

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Main Authors: Hanapi, Ummi Kalthum, Mohd Desa, Mohd Nasir, Ismail, Amin, Mustafa, Shuhaimi
Format: Article
Language:English
Published: Springer 2015
Online Access:http://psasir.upm.edu.my/id/eprint/36170/1/A%20higher%20sensitivity%20and%20efficiency%20of%20common%20primer%20multiplex%20PCR%20assay%20in%20identification%20of%20meat%20origin%20using%20NADH%20dehydrogenase%20subunit%204%20gene.pdf
http://psasir.upm.edu.my/id/eprint/36170/
http://link.springer.com/article/10.1007%2Fs13197-014-1459-7
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spelling my.upm.eprints.361702015-09-16T23:29:23Z http://psasir.upm.edu.my/id/eprint/36170/ A higher sensitivity and efficiency of common primer multiplex PCR assay in identification of meat origin using NADH dehydrogenase subunit 4 gene Hanapi, Ummi Kalthum Mohd Desa, Mohd Nasir Ismail, Amin Mustafa, Shuhaimi A Common Primer Multiplex PCR (CP-M-PCR) was developed to detect meat origin of four groups of animal (pig, ruminant, avian and rabbit). This method demonstrated higher sensitivity and efficiency than the conventional multiplex PCR. In this approach, a common forward primer was designed in the 5′ end of a homologous region of mitochondrial NADH dehyrogenase subunit 4 (Nad 4) gene sequences of all the animal groups. Specific adapter reverse primers were designed by adding an adapter sequence at the 5′ end. The same adapter sequence was used as the common adapter reverse primer. The primers generated specific fragments of 267, 370, 504, and 548 bp lengths for pig, ruminant, avian and rabbit meats, respectively. The use of adapter sequence at the 5′ end of the common adapter reverse primers increased the efficiency of the amplification and the application of a common forward primer solved the complexity in multiplex PCR system. Bands of specific amplification can be detected in the PCR assays containing as low as 10−6 μM of adapter reverse primer. This result indicated that the sensitivity was tremendously increased as compared to the conventional multiplex PCR (10−3 μM). CP-M-PCR detection limit of the DNA samples was 0.1 ng for the four groups of meats. CP-M-PCR has greatly improved the sensitivity and efficiency of the PCR system for a more reliable and accurate outcome than conventional multiplex PCR system. Springer 2015 Article PeerReviewed application/pdf en http://psasir.upm.edu.my/id/eprint/36170/1/A%20higher%20sensitivity%20and%20efficiency%20of%20common%20primer%20multiplex%20PCR%20assay%20in%20identification%20of%20meat%20origin%20using%20NADH%20dehydrogenase%20subunit%204%20gene.pdf Hanapi, Ummi Kalthum and Mohd Desa, Mohd Nasir and Ismail, Amin and Mustafa, Shuhaimi (2015) A higher sensitivity and efficiency of common primer multiplex PCR assay in identification of meat origin using NADH dehydrogenase subunit 4 gene. Journal of Food Science and Technology, 52 (7). pp. 4166-4175. ISSN 0022-1155; ESSN: 0975-8402 http://link.springer.com/article/10.1007%2Fs13197-014-1459-7 10.1007/s13197-014-1459-7
institution Universiti Putra Malaysia
building UPM Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Putra Malaysia
content_source UPM Institutional Repository
url_provider http://psasir.upm.edu.my/
language English
description A Common Primer Multiplex PCR (CP-M-PCR) was developed to detect meat origin of four groups of animal (pig, ruminant, avian and rabbit). This method demonstrated higher sensitivity and efficiency than the conventional multiplex PCR. In this approach, a common forward primer was designed in the 5′ end of a homologous region of mitochondrial NADH dehyrogenase subunit 4 (Nad 4) gene sequences of all the animal groups. Specific adapter reverse primers were designed by adding an adapter sequence at the 5′ end. The same adapter sequence was used as the common adapter reverse primer. The primers generated specific fragments of 267, 370, 504, and 548 bp lengths for pig, ruminant, avian and rabbit meats, respectively. The use of adapter sequence at the 5′ end of the common adapter reverse primers increased the efficiency of the amplification and the application of a common forward primer solved the complexity in multiplex PCR system. Bands of specific amplification can be detected in the PCR assays containing as low as 10−6 μM of adapter reverse primer. This result indicated that the sensitivity was tremendously increased as compared to the conventional multiplex PCR (10−3 μM). CP-M-PCR detection limit of the DNA samples was 0.1 ng for the four groups of meats. CP-M-PCR has greatly improved the sensitivity and efficiency of the PCR system for a more reliable and accurate outcome than conventional multiplex PCR system.
format Article
author Hanapi, Ummi Kalthum
Mohd Desa, Mohd Nasir
Ismail, Amin
Mustafa, Shuhaimi
spellingShingle Hanapi, Ummi Kalthum
Mohd Desa, Mohd Nasir
Ismail, Amin
Mustafa, Shuhaimi
A higher sensitivity and efficiency of common primer multiplex PCR assay in identification of meat origin using NADH dehydrogenase subunit 4 gene
author_facet Hanapi, Ummi Kalthum
Mohd Desa, Mohd Nasir
Ismail, Amin
Mustafa, Shuhaimi
author_sort Hanapi, Ummi Kalthum
title A higher sensitivity and efficiency of common primer multiplex PCR assay in identification of meat origin using NADH dehydrogenase subunit 4 gene
title_short A higher sensitivity and efficiency of common primer multiplex PCR assay in identification of meat origin using NADH dehydrogenase subunit 4 gene
title_full A higher sensitivity and efficiency of common primer multiplex PCR assay in identification of meat origin using NADH dehydrogenase subunit 4 gene
title_fullStr A higher sensitivity and efficiency of common primer multiplex PCR assay in identification of meat origin using NADH dehydrogenase subunit 4 gene
title_full_unstemmed A higher sensitivity and efficiency of common primer multiplex PCR assay in identification of meat origin using NADH dehydrogenase subunit 4 gene
title_sort higher sensitivity and efficiency of common primer multiplex pcr assay in identification of meat origin using nadh dehydrogenase subunit 4 gene
publisher Springer
publishDate 2015
url http://psasir.upm.edu.my/id/eprint/36170/1/A%20higher%20sensitivity%20and%20efficiency%20of%20common%20primer%20multiplex%20PCR%20assay%20in%20identification%20of%20meat%20origin%20using%20NADH%20dehydrogenase%20subunit%204%20gene.pdf
http://psasir.upm.edu.my/id/eprint/36170/
http://link.springer.com/article/10.1007%2Fs13197-014-1459-7
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score 13.211869

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