Rapid detection of porcine DNA in processed food samples using a streamlined DNA extraction method combined with the SYBR Green real-time PCR assay

Tan, Lee Lee; Ahmed, Siti Aminah; Ng, Siew Kit; Citartan, Marimuthu; Raabe, Carsten A.; Rozhdestvensky, Timofey S.; Tang, Thean‐Hock

doi:10.1016/j.foodchem.2019.125654

Cited by 14 publications

(5 citation statements)

References 29 publications

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“…In this work, different types of dry and wet matrixes were analysed (kibble, pâté, bites with sauce, and strips), all of which are the result of technological processes such as mechanical, thermal, chemical and enzymatic treatments. In such cases, specific commercial DNA extraction kits and customized DNA extraction protocols 30,40 are required to ensure the isolation of high-quality DNA. In this study, the protocol described by Lagisz et al 41 was tested, resulting in good efficiency in terms of DNA quantity and the 260/280 ratio.…”

Section: Discussionmentioning

confidence: 99%

NGS-based barcoding with mini-COI gene target is useful for pet food market surveys aimed at mislabelling detection

Palumbo

Scariolo

Vannozzi

et al. 2020

Sci Rep

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Pet food industry has grown considerably in the last few years and it is expected to continue with this rate. Despite the economic impact of this sector and the consumer concerns for the increasing number of food and feed adulteration cases, few studies have been published on mislabelling in pet foods. We therefore investigated the capability of a next generation sequencing-based mini-barcoding approach to identify animal species in pet food products. In a preliminary analysis, a 127 bp fragment of the COI gene was tested on both individual specimens and ad hoc mixed fresh samples used as testers, to evaluate its discrimination power and primers effectiveness. Eighteen pet food products of different price categories and forms available on the market (i.e. kibbles, bites, pâté and strips) were analysed through an NGS approach in biological replicates. At least one of the species listed in the ingredients was not detected in half of the products, while seven products showed supplementary species in addition to those stated on the label. Due to the accuracy, sensitivity and specificity demonstrated, this method can be proposed as food genetic traceability system to evaluate both the feed and food quality timely along the supply chain.

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Section: Discussionmentioning

confidence: 99%

NGS-based barcoding with mini-COI gene target is useful for pet food market surveys aimed at mislabelling detection

Palumbo

Scariolo

Vannozzi

et al. 2020

Sci Rep

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“…Probe mempunyai kelebihan seperti multiplek target dalam satu sumur, konsumsi waktu yang cepat, dan sensitivitas tinggi (Kim & Kim, 2019). Probe qPCR telah berhasil digunakan untuk mendeteksi cemaran DNA babi di produk pangan, seperti roti, bakso, dan permen, serta produk nonpangan, seperti kuas roti, pelembap, sabun, kosmetik, dan kapsul (Kim & Kim, 2019;Kim et al, 2018;Mohamad et al, 2018;Mustaqimah et al, 2021;Septiani, 2021;Sudjadi et al, 2016;Tan et al, 2020;Tanabe et al, 2007;Widayat et al, 2019).…”

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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“…Stronger signals for the raw anchovy presumably are resulted by higher relative binding of SYBR Green to the longer fragment relatively shorter anchovy -specific amplicons which cause to higher melting peak (Marín et al, 2010). Lee Tan et al (2019) also highlighted that the thermally processing lead to variance in melting curve.…”

Section: Differences In Dna Degradationmentioning

confidence: 99%

Effect of Industrial and Domestic Processing Techniques On the DNA Degradation of Anchovy

Tümerkan

2022

Turkish JAF Sci.Tech.

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Food fraudulent activities have become a serious issue over the world recently. Seafood products have trading and profitable potential in Turkey owing to the abundance of fisheries and other species. While morphological features are commonly used for species identification in raw seafood products, this identification does not meet the correct classification in cryptic species and processed seafood products. Molecular techniques have been utilized for species authentication in processed seafood items successfully. In this study, the effect of different processing techniques on the DNA quality and DNA degradation isolated from raw and processed anchovy was investigated. Anchovy is one of the important species in both fisheries activities and processing and consumption in Turkey. For this aim, DNA was isolated from processed anchovy groups and un-processed anchovy groups as control by the same extraction methods and the quality of DNA was compared among the groups. The most common processing techniques, frying, baking, smoking, roasting, baking and grilling were applied to anchovy. The results revealed that not only different thermal processing but also treatment with acid and salt cause DNA degradation and quality loss of DNA parameters which are essential for authentication of species and traceability for public health.

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Rapid detection of porcine DNA in processed food samples using a streamlined DNA extraction method combined with the SYBR Green real-time PCR assay

Cited by 14 publications

References 29 publications

NGS-based barcoding with mini-COI gene target is useful for pet food market surveys aimed at mislabelling detection

NGS-based barcoding with mini-COI gene target is useful for pet food market surveys aimed at mislabelling detection

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

Effect of Industrial and Domestic Processing Techniques On the DNA Degradation of Anchovy

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