Determination of Porcine Contamination in Laboratory PrepareddendengUsing Mitochondrial D-Loop686 andcyt bGene Primers by Real Time Polymerase Chain Reaction

Abstract

The identifications of species in meat products have created interests since these foods became the target of forgery and fraud in the market. The presence of pork in food products is not allowed for the Muslim community. Hence, an analysis is necessary to detect the presence of pork in processed meat products, such as in dendeng (dried meat) product. Real time polymerase chain reaction using mitochondrial displacement loop686 and cytochrome b (cytb) gene primers was used to identify specific pork DNA among other four types of DNA species; namely beef, chicken, goat, and horse. This method was also used to identify pork DNA in the laboratory processed pork-beef dendeng as well as commercial dendeng from market. The results showed that real time polymerase chain reaction using displacement loop686 and cytb gene primers were able specifically to distinguish between pork DNA and the other species. The lowest concentration of 0.5% of pork DNA in a mixture of pork-beef processed products of dendeng was able to be detected by both primers with the product amplification of 114 and 134 bp (base pair) for the displacement loop686 and 149 bp for cytb gene, respectively. High sensitivity was also obtained when both primers were applied with the lowest detection limit of 5 pg/µL pork DNA. The results of the six commercial dendeng amplification using both primers showed no amplified products present, meaning that these products do not contain porcine DNA.

Molecular Based Method Using PCR Technology on Porcine Derivative Detection for Halal Authentication

“…The cytb gene primers also had a similar ability with d-Loop686 primer to amplify pork DNA until the pork concentration was of 0.5% in beef "dendeng". The results showed that these two primers, d-Loop region and the cyt b gene, had the capability to identify pork adulteration on the mixed pork-beef "dendeng" product at all variations of tested concentrations [42].…”

“…Animal Species PCR assay for the identification of animal species in feedstuffs [51] Analysis of rat's meat DNA Authentication of beef meatball from rat's meat [52] Analysis of pork DNA in dendeng Authentication of "dendeng" from pork [42] Analysis of porcine gelatin Analysis of porcine gelatin in capsule shell [53] Analysis of pork in meatball Authentication of beef meatball from pork [34] Analysis of pork DNA Identification of pig species in food product [1] Quantification of bovine, porcine, chicken and turkey species Contamination and adulteration prohibited component in food and feed [54] Porcine Gelatin Using gelatin in pharmacy and confectionery [55] Table 3. Analysis of porcine species-based PCR technology on commercial samples.…”

“…The previous research [42] performed the real-time polymerase chain reaction using mitochondrial displacement loop686 and cytochrome b (cytb) gene primers for the identification specific pork DNA among other four types of DNA species: beef, chicken, goat and horse. The annealing temperature was at 62°C; however, the mitochondrial D-Loop686 showed high specificity to porcine DNA detection, as indicated by no amplification results that appeared on other species (beef, chicken, horse and goat meat) ( Figure 5).…”

Application method of Fourier Transform Infrared (FTIR) combined with chemometrics for analysis of rat meat (Rattus Diardi) in meatballs beef

“…Dengan kemajuan ilmu pengetahuan dan teknologi modern, mulai ada produk makanan yang dicampur dengan bahan non-halal (Van der Spiegel et al, 2012;Schieber, 2008). Sebagian besar komponen non-halal yang ditemukan di pasar adalah daging babi, lemak babi, gelatin babi, dan produk-produk berbasis babi (Maryam et al, 2016;Rahmawati et al, 2016;Sudjadi et al, 2016). Bahan-bahan yang terdapat dalam produk makanan, farmasi atau produk kosmetika dapat bersifat diperbolehkan untuk dikonsumsi secara hukum Islam (halal) atau dilarang dikonsumsi menurut hukum Islam (haram) (Mursyidi, 2013;Raraswati et al, 2013).…”

Forgery Detection Beef with Mice Meat (<i>Mus musculus</i>) in Meatballs Using Real-Time Polymerase Chain Reaction (Real-Time PCR) Primer Specific for a Target Mitochondrial DNA ND-1 Gene

“…There are several types of PCR including PCR-RAPD, PCR-RFLP and PCR primer specific and real-time PCR. These techniques have been reported the presence of porcine in food including PCR primer specific [3][4]; PCR-RFLP [5][6], PCR-RAPD [7][8], real-time PCR [9][10]. In case of detection of mice, only a few work has been reported.…”