Real Time PCR Assays for Detection and Quantification of Porcine DNA in Meat Milling Samples

Baihaqi, M.; Rachmawati, Yuanita; Rokhim, Saiku; Munir, Misbakhul; Hamidah, Lilik

doi:10.4108/eai.2-5-2019.2284634

Cited by 3 publications

(3 citation statements)

References 15 publications

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“…Jaminan produk halal pada pangan agar terhindar dari cemaran babi berdasarkan pada risiko mengonsumsi daging babi, seperti tingginya lemak meningkatkan obesitas, diabetes tipe 2, dan menyebabkan alergi (Bott, 2014;Rupa et al, 2009). Cemaran babi terdeteksi di berbagai produk makan, seperti mesin penggilingan daging, daging giling, bakso, gelatin, dan sosis (Baihaqi et al, 2019;Laila-Liyana et al, 2018).…”

Section: Pendahuluanunclassified

Deteksi dan Kuantifikasi Cemaran Babi pada Sampel Olahan Daging Menggunakan Real-time PCR

Waluyo

Malau

Raekiansyah

et al. 2023

Al-Kauniyah J. Biol.

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AbstrakMetode pengujian cemaran babi menjadi faktor penting dalam sertifikasi produk halal. Metode yang cepat dan robust diperlukan untuk deteksi dan kuantifikasi cemaran babi. Metode Real-time PCR atau dikenal dengan istilah quantitative PCR (qPCR) merupakan metode alternatif untuk deteksi dan kuantifikasi cemaran babi berdasarkan residu keberadaan DNAnya pada sampel olahan pangan. Metode ekstraksi DNA dan kit amplifikasi yang tahan terhadap inhibitor menjadi kunci keberhasilan penggunaan qPCR untuk pendeteksian dan kuantifikasi cemaran babi. Pendeteksian cemaran DNA dengan probe qPCR digunakan karena mempunyai kelebihan tahan terhadap inhibitor, cepat, spesifik, dan multipel target. Penelitian ini bertujuan untuk mendeteksi dan menguantifikasi cemaran DNA babi menggunakan metode ekstraksi DNA secara cepat dan qPCR. Tahapan penelitian ini adalah ekstraksi DNA, amplifikasi, deteksi, dan kuantifikasi DNA babi. Sampel berasal dari produk olahan pangan, seperti bakso, sosis, daging burger, siomay, kuah daging, dan daging isi roti. Hasil penelitian menunjukkan bahwa terdapat cemaran babi pada sampel bakso, daging burger, dan kuah bakso. Hasil yang didapatkan menunjukkan bakso memiliki persentase kontaminasi sejumlah 25%, sedangkan kuah daging sejumlah 12,5%. Hasil penelitian ini dapat direkomendasikan untuk laboratorium penguji makanan sebagai metode deteksi cemaran babi dalam produk pangan secara cepat dan akurat.AbstractPork contamination testing method is an important factor in halal product certification. A fast and robust method is needed for the detection and quantification of pig contamination. Real-time PCR method or commonly known as quantitative PCR (qPCR) is an alternative method for the detection and quantification of pork contamination based on the pig’s DNA residual presence in processed food samples. DNA extraction method and inhibitor-resistant amplification kit are the keys of successful qPCR implementation for the detection and quantification of pig contamination. Detection of DNA contamination with qPCR probe is used because it has some advantages, such as resistant to inhibitors, fast, specific, and multiple targets. This research aimed to detect and quantify pig’s DNA contamination using rapid DNA extraction method and qPCR. The stages of this research were pig’s DNA extraction, amplification, detection, and quantification. The samples taken from processed food products, such as meatballs, sausage, burgers’ meat, dumplings, meat broth, and meat filled in the bread. The results showed that there was pork contamination in the samples of meatballs, burgers’ meat, and meat broth. The results showed that the meatballs had a contamination percentage of 25%, while the meat broth had a contamination percentage of 12.5%. The results of this study can be a recommendation for food testing laboratories as a method of detecting the pork contamination in food products quickly and accurately.

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Section: Pendahuluanunclassified

Deteksi dan Kuantifikasi Cemaran Babi pada Sampel Olahan Daging Menggunakan Real-time PCR

Waluyo

Malau

Raekiansyah

et al. 2023

Al-Kauniyah J. Biol.

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“…According to Baihaqi et al (2019) some of food produced by micro-entrepreneurs still uses general meat grinding in market which can cause mixing of various types of meat from other customers. Products that have been given a mixture of pork are very difficult to differentiated, because the meat is mashed and then mixed with other ingredients.…”

Section: Introductionmentioning

confidence: 99%

End Point Polymerase Chain Reaction for Porcine Detection on Food Product of UIN Sunan Ampel Surabaya Canteen

Rokhim¹,

Tyautari²,

Firmansyah³

et al. 2021

Indonesian Journal of Halal Research

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Halal food means food that permitted under Islamic law and fulfills about requirements. The absence of information about halal food contained in UIN Sunan Ampel Surabaya (UINSA) campus area causes related research to be carried out. This study aims to determine the porcine DNA contamination on food around UINSA area using molecular technology. Twenty two samples used were foods that contain meat and may contain pork obtained from canteens around UINSA area, analyzed using Polymerase Chain Reaction (PCR) method. The analysis was started with DNA isolation of 22 food samples, electrophoresis, PCR, then visualization gel electrophoresis. Primer gene coding for cytochrome b (cyt b) which produces 149 bp of DNA fragments. The results showed that no porcine contamination in 22 food samples, while the positive control showed a band of 149 bp. End point PCR method potentially to detect porcine DNA contaminants in food products around UINSA. Therefore the food is halal and safe for consumption.

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“…Micro entrepreneurs with basic ingredients such as processed meat who do not have a grinding tool, will decide grind meat to a public meat grinding in the market (Baihaqi, Rachmawati, Rokhim, Munir, & Hamidah, 2019). Product counterfeiting is a problem that often occurs given the significant increase in profits and to reduce production costs.…”

Section: Introductionmentioning

confidence: 99%

Sus sp. DNA Encoding cyt b Gene Detection Test on Meat Grinding Samples Using Conventional PCR

Adzakiyyi

Susilowati

Rokhim

et al. 2020

Indonesian Journal of Halal Research

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Micro-entrepreneurs with basic ingredients of processed meat such as meatball who do not have a meat grinder, generally using meat grinder at the public market. The problem that occurs is that there is no clear regulation from the Government regarding the guarantee of the halal meat grinding in the Regional Company. This needs to be enhanced as a study, considering that the grinding material does not only come from Halal substances. The purpose of this study was to test pig DNA in meat grinding samples at PD Pasar Surya Surabaya City by using the conventional Polymerase Chain Reaction (PCR) method. DNA was isolated from 11 PD Pasar Surya meat grinding samples, then spectrophotometry was performed. Spectrophotometry results showed that all samples have high DNA concentrations. The primer used is the cyt b pig gene encoder. Predenaturation is performed at a temperature of 95°C-5 minutes, denaturation of 95°C-45 seconds, annealing 60°C-30 seconds, extension 72°C-40 seconds, and post extension 72°C-5 minutes. The results of PCR analysis were determined by the emergence of DNA bands of ± 149 bp as markers of pig DNA. The results showed negative on sample or no pig contamination in 11 samples tested. While the pig sample as positive control showed a band of ± 149 bp. These results prove that at 11 points of the location of meat grinding there is no contamination of pig DNA.

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Real Time PCR Assays for Detection and Quantification of Porcine DNA in Meat Milling Samples

Cited by 3 publications

References 15 publications

Deteksi dan Kuantifikasi Cemaran Babi pada Sampel Olahan Daging Menggunakan Real-time PCR

Deteksi dan Kuantifikasi Cemaran Babi pada Sampel Olahan Daging Menggunakan Real-time PCR

End Point Polymerase Chain Reaction for Porcine Detection on Food Product of UIN Sunan Ampel Surabaya Canteen

Sus sp. DNA Encoding cyt b Gene Detection Test on Meat Grinding Samples Using Conventional PCR

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