Article

Comparison of seven commercial TaqMan master mixes and two real-time PCR platforms regarding the rapid detection of porcine DNA

Soo Ji Kang1, Chan Song Jang1, Ji Min Son1, Kwang Won Hong1,*
Author Information & Copyright
1Dongguk University, Ilsandong-gu, Goyang-si 10326, Korea.
*Corresponding Author: Kwang Won Hong, Dongguk University, Ilsandong-gu, Goyang-si 10326, Korea, Republic of. Phone: +82-31-961-5140. E-mail: hkwon@dongguk.edu.

© Copyright 2020 Korean Society for Food Science of Animal Resources. This is an Open-Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Received: Jun 08, 2020 ; Revised: Aug 19, 2020 ; Accepted: Sep 11, 2020

Published Online: Sep 16, 2020

Abstract

A pig-specific real-time PCR assay based on the mitochondrial ND5 gene was developed to detect porcine material in food and other products. To optimize the performance of assay, seven commercial TaqMan master mixes and two real-time PCR platforms (Applied Biosystems StepOnePlus and Bio-rad CFX Connect) were used to evaluate the limit of detection (LOD) as well as the PCR efficiency and specificity. The LODs and PCR efficiencies for the seven master mixes on two platforms were 0.5–5 pg/reaction and 84.96%–108.80%, respectively. Additionally, non-specific amplifications of DNA from other animal samples (human, dog, cow, and chicken) were observed for four master mixes. These results imply that the sensitivity and specificity of a real-time PCR assay may vary depending on master mix and platform used. The best combination of master mix and real-time PCR platform can accurately detect 0.5 pg porcine DNA, with a PCR efficiency of 100.49%.

Keywords: master mix; real-time PCR; species identification; porcine DNA


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The name of the journal has been changed from Korean Journal for Food Science of Animal Resources to Food Science of Animal Resources from January 2019.

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