A quick and simple method for the identification of meat species and meat products by PCR assay

Matsunaga, Takateru; Chikuni, Koichi; Tanabe, Ryo; Muroya, Susumu; Shibata, Kiyohiro; Yamada, Junji; Shinmura, Yutaka

doi:10.1016/s0309-1740(98)00112-0

Cited by 365 publications

(283 citation statements)

References 9 publications

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“…Amplification was conducted on a cytochrome b gene with a common forward primer and specific reverse primers, as first described previous literature [18]. The size calculated from these samples were different than previously reported [13], whereas bovine samples, a band size of 320 bp was calculated, and from porcine samples a band size of 360 bp was calculated.…”

Section: Pcr Amplification Showed Varied Results Depending On Dna Isomentioning

confidence: 99%

Comparison of DNA Extraction Methods Between Conventional, Kit, Alkali and Buffer-Only for PCR Amplification on Raw and Boiled Bovine and Porcine Meat

Yahya

Firmansyah

Arlisyah³

et al. 2017

JELS

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Detection of porcine contamination in food material by employing PCR techniques is integral in halal food confirmation. However, PCR is both costly and laborious, particularly in DNA isolation method. This study explores several different methods in DNA extraction for PCR amplification in bovine and porcine raw and boiled meat samples. Four methods for DNA extraction (conventional PCI method, DNA isolation kit, alkaline-based method, and a DNA lysis buffer-only from the same kit) was employed followed by PCR using primers from previous studies and compared for DNA quality and quantity (in six replicates) and PCR amplification on the best three DNA samples. This study shows that in all samples, the conventional method had the best DNA yield based on nanodrop measurement, followed by an alkali-based method, buffer-only method, and DNA isolation kit. Each method except lysis-buffer only had at least one sample with good DNA quality. Conventional and isolation kit showed reliable positive PCR detection for all porcine and bovine samples (92% positive). Using the alkaline-lysis method, DNA was amplified reliably on boiled meat samples (83% positive). Lysis-buffer-only method did not show consistent PCR amplification on the samples used (50% positive). The conclusion was that conventional PCI method and DNA isolation kit showed high reliability in PCR amplification of bovine and porcine meats, both raw and boiled. While high DNA yield was obtained using the alkaline-lysis method, PCR amplification was only successful on boiled samples. Lysis-buffer only method yielded in poor DNA quality and was not able to result in reliable DNA amplification.

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Section: Pcr Amplification Showed Varied Results Depending On Dna Isomentioning

confidence: 99%

Comparison of DNA Extraction Methods Between Conventional, Kit, Alkali and Buffer-Only for PCR Amplification on Raw and Boiled Bovine and Porcine Meat

Yahya

Firmansyah

Arlisyah³

et al. 2017

JELS

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“…The PCR was carried out to amplify fragments of DNA mitochondrial Cyt-b gene, and the primer used in this study was previously published by Matsunaga et al (1999) which is presented in Table 1. PCR program was performed using the GeneAmp® PCR System 9700 (Singapore) as follows: initial temperature of denaturation at 95 o C for 3 minutes, followed by 30 cycles of denaturation at 95 o C for 15 seconds, annealing temperature at 60 o C for 30 seconds and extension at 72 o C for 30 seconds and also final extension at 72 o C for 1 minute.…”

Section: Simplex-and Duplex-pcrmentioning

confidence: 99%

Autentikasi Daging Ayam Segar Dari Kontaminasi Daging Babi Menggunakan Gen Cyt-B Dengan Analisis Duplex- Polymerase Chain Reaction

Hertanto¹,

Fitra²,

Kartikasari³

et al. 2017

BuletinPeternak

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Halal is one of important aspects in consumer protection. Meat and processed meat products are food that should be controlled strictly because those are prone to be adulterated by pork contamination. Therefore, it is necessary to provide detection technique which is accurate, fast and cheap. The objective of this research was to identify the presence of impurities of pork meat on raw chicken meat using gene Cyt-b with duplex-PCR analysis. This research used six samples of raw chicken meat and raw pork. Raw chicken meat was bought from supermarkets in the city of Surakarta and raw pork was obtained from pig slaughterhouse. The percentage of raw pork contamination on raw chicken meat was designed as much as 1, 5, 10, and 25%, respectively. The DNA genome was isolated according to DNA isolation protocol from Genomic DNA Mini Kit. In addition, duplex-PCR was performed based on protocol of KAPA2G Fast Multiplex PCR kit. The data was descriptively analyzed by directly looking the DNA bands on the gel documentation apparatus. The result showed that specific DNA bands for chicken and pig were completely appeared on 1.5% of agarose gels. Duplex-PCR detect contamination of pork on raw meat of chicken at all contamination levels. This research proved that the duplex-PCR detect the contamination of pork until the level of 1%.

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“…Polymerase Chain Reaction (PCR) technique was reported very precise to detect meat adulteration in meat species identification for routine analysis (Matsunaga et al 2001). The use of seven restriction enzyme endonucleases in this PCR-Restriction Fragment Length Polymorphism (PCR-RFLP) analysis were expected to generate a unique genetic profiling for each meat species, based on the size and number of DNA fragments being produced after the enzymatic digestion of cyt b amplicons.…”

Section: Introductionmentioning

confidence: 99%

Comparison of DNA Profiling between Fishes and Pork Meat using Polymerase Chain Reaction-Restriction Fragment Length Polymorphisms (PCR-RFLP) Analysis

Shahimi

Mutalib²,

Nazri

et al. 2018

JSM

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A quick and simple method for the identification of meat species and meat products by PCR assay

Cited by 365 publications

References 9 publications

Comparison of DNA Extraction Methods Between Conventional, Kit, Alkali and Buffer-Only for PCR Amplification on Raw and Boiled Bovine and Porcine Meat

Comparison of DNA Extraction Methods Between Conventional, Kit, Alkali and Buffer-Only for PCR Amplification on Raw and Boiled Bovine and Porcine Meat

Autentikasi Daging Ayam Segar Dari Kontaminasi Daging Babi Menggunakan Gen Cyt-B Dengan Analisis Duplex- Polymerase Chain Reaction

Comparison of DNA Profiling between Fishes and Pork Meat using Polymerase Chain Reaction-Restriction Fragment Length Polymorphisms (PCR-RFLP) Analysis

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