Detection and quantification of pork and rat DNA in processed meats using multiplex quantitative Real‐Time PCR (m‐qPCR)

Nurul Azmah Nikmatullah; Etin Diah Permanasari

doi:10.22146/ijbiotech.94212

Detection and quantification of pork and rat DNA in processed meats using multiplex quantitative Real‐Time PCR (m‐qPCR)

https://doi.org/10.22146/ijbiotech.94212

Nurul Azmah Nikmatullah⁽¹⁾, Etin Diah Permanasari^(2*)

(1) Health Analyst Study Program, Faculty of Pharmacy, University of Muhammadiyah Prof. DR. HAMKA, East Jakarta, DKI Jakarta 13460, Indonesia; Center for Halal Studies, University of Muhammadiyah Prof. DR. HAMKA, East Jakarta, DKI Jakarta 13460, Indonesia
(2) Master of Pharmaceutical Science, Postgraduate School, University of Muhammadiyah Prof. DR. HAMKA, South Jakarta, DKI Jakarta 12740, Indonesia; Center for Halal Studies, University of Muhammadiyah Prof. DR. HAMKA, East Jakarta, DKI Jakarta 13460, Indonesia
(*) Corresponding Author

Abstract

In addition to the issue of pork contamination, processed meats frequently contain traces of rat meat. Therefore, detection and quantification of the pork and rat DNA in cases of meat and processed meat adulteration are necessary. In the current study, two gene targets of the cytochrome b for pigs and the Mt‐atp6 of Rattus norvegicus for rats were used in the absolute multiplex quantitative real‐time PCR (m‐qPCR). The sample DNA was amplified with a standard as positive control in the various concentration of 1000 pg, 100 pg, 10 pg, 0.1 pg, 0.01 pg, and 0.001 pg. There were 25 processed meat samples and 5 fresh meat samples identified in this study. Among the total of 30 samples assessed, 6 samples were successfully detected and quantified their pork and rat DNA contamination. One sample was contaminated with pork DNA with a concentration of 2.451×10^‐4pg (“Meatball 3). Five samples were contaminated with rat DNA with a concentration of 3.603×10^‐11pg (“Sempol 3”), 2.196×10^‐10pg (“Meatball 6”), 4.908×10^‐11pg (“Siomay 3”), 1.489×10^‐10pg (“Grinding 2”), and 3.564×10^‐10pg (“Grinding 4”). In this study, we have discovered that the contamination of pork and rat were detected in the samples. It suggested that this method is applicable for detecting the contaminant in processed meat samples

Keywords

Cytochrome b; Multiplex PCR; Mt‐atp6; Processed meat; qPCR

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DOI: https://doi.org/10.22146/ijbiotech.94212