Species Authentication of Dog, Cat, and Tiger Using Cytochrome β Gene

Irine,; Nuraini, Henny; Sumantri, Cece

doi:10.5398/medpet.2013.36.3.171

Cited by 7 publications

(7 citation statements)

References 23 publications

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“…Previous studies successfully identified various species in the meats with and without temperature treatments by using multiplex PCR mitochondrial DNA cytochrome b gene (Matsunaga et al, 1999;Shin et al, 2006;Ni'mah et al, 2016;Hertanto et al, 2017). Irine et al (2013) also used cytochrome b gene to identify species of tiger, cat, and dog. In addition to cytochrome b gene, another unique region contained in animal mitochondrial genome is 12S rRNA gene.…”

Section: Introductionmentioning

confidence: 99%

Development of mitochondrial 12S rRNA gene for identification of dog and rat in beef using multiplex PCR

Cahyadi

Taufik

Pramono

et al. 2019

J. Indonesian Trop. Anim. Agric.

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The 12S rRNA gene is one of unique regions in mitochondrial genome usually used for phylogenetic studies and species identification. The objective of present study was to develop species specific primers from mitochondrial 12S rRNA gene for identification of dog and rat in beef by using multiplex PCR assay. Three primer pairs of mitochondrial 12S rRNA gene specific for bovine, dog and rat were designed and selected to evaluate their specificity and fidelity. Moreover, a total of twelve DNA samples extracted from meat tissue were also prepared to test those primers using simplex and multiplex PCR. The PCR products were then visualized using 2% of agarose gel under the UV light and three of them were sequenced. In addition, sequence data were analyzed using Clustal Omega software and BLAST. The result showed that simplex PCR assay successfully amplified DNA targets which are respectively indicated by 155 bp (bovine), 244 bp (dog), and 491 bp (rat) of DNA bands. Furthermore, DNA sample sequences were identically similar to reference sequence used in this study. Multiplex and simplex PCR analyses also indicated that these primer pairs specifically amplified DNA target for each species in the samples containing various species. The results suggested that designed primers in this study could be used to identify dog and rat in raw beef containing these species meat. Further experiment should be conducted using meat-processed products and commercial meat products as samples.

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Section: Introductionmentioning

confidence: 99%

Development of mitochondrial 12S rRNA gene for identification of dog and rat in beef using multiplex PCR

Cahyadi

Taufik

Pramono

et al. 2019

J. Indonesian Trop. Anim. Agric.

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“…Previously developed feline-specific PCR assays were based on mitochondrial whole genome: 672 bp (Abdulmawjood et al 2003); 672 bp (Abdel- Rahman et al 2009); ND4 gene (274 bp) (Ilhak & Arslan 2007); 12S rRNA (108 bp) (Martin et al 2007); cytb gene (180 bp) (Tobe & Linacre 2008); and cytb gene (331 bp) (Irine et al 2013). These assays involved considerably longer amplicons which are assumed to be fragmented under commercial food processing treatments, causing PCR failure or truncated PCR products.…”

Section: Results and Discussion Feline Species Specificitymentioning

confidence: 99%

“…To study the comparative stability of the newly designed 69 bp amplicon and one of the previously documented 108 bp targets (the shortest of the previously documented targets 672 (Abdel- Rahman et al 2009), 331 (Irine et al 2013), 274 (Ilhak & Arslan 2007), 180 (Tobe & Linacre 2008) and 108 bp (Martin et al 2007)). PCR was performed with a 50 ng feline DNA template after autoclaving feline meats at 135°C for 30, 60, 90, 120 and 150 min under 0.3 MPa and microwave cooking at 600 and 700 W for 30 min.…”

Section: Comparison Of Target Dna Stabilitymentioning

confidence: 99%

Lab-on-a-chip-based PCR-RFLP assay for the confirmed detection of short-length feline DNA in food

Ali

Amin

et al. 2015

Food Additives & Contaminants: Part A

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Wider availability but lack of legal market trades has given feline meat a high potential for use as an adulterant in common meat and meat products. However, mixing of feline meat or its derivatives in food is a sensitive issue, since it is a taboo in most countries and prohibited in certain religions such as Islam and Judaism. Cat meat also has potential for contamination with of severe acute respiratory syndrome, anthrax and hepatitis, and its consumption might lead to an allergic reaction. We developed a very short-amplicon-length (69 bp) PCR assay, authenticated the amplified PCR products by AluI-restriction digestion followed by its separation and detection on a lab-on-a-chip-based automated electrophoretic system, and proved its superiority over the existing long-amplicon-based assays. Although it has been assumed that longer DNA targets are susceptible to breakdown under compromised states, scientific evidence for this hypothesis has been rarely documented. Strong evidence showed that shorter targets are more stable than the longer ones. We confirmed feline-specificity by cross-challenging the primers against 10 different species of terrestrial, aquatic and plant origins in the presence of a 141-bp site of an 18S rRNA gene as a universal eukaryotic control. RFLP analysis separated 43- and 26-bp fragments of AluI-digest in both the gel-image and electropherograms, confirming the original products. The tested detection limit was 0.01% (w/w) feline meat in binary and ternary admixed as well as meatball matrices. Shorter target, better stability and higher sensitivity mean such an assay would be valid for feline identification even in degraded specimens.

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“…Previously, cat meat was detected by PCR assay targeting a 672-bp (Abdulmawjood et al 2003;Abdel Rahman et al 2009); 274-bp (ND4) (İlhak and Arslan 2007); 108-bp (12S ribosomal RNA (rRNA)) (Martín et al 2007); 180-bp (Tobe and Linacre 2008), and 331-bp sites of cytb gene (Irine et al 2013). However, it has been assumed that longer amplicons are thermodynamically less stable than the shorter ones under environmental decomposition or degradation by food processing treatment Ali et al 2011;Rahman et al 2014).…”

Section: Specificity Testmentioning

confidence: 99%

Short Amplicon-Length PCR Assay Targeting Mitochondrial Cytochrome b Gene for the Detection of Feline Meats in Burger Formulation

et al. 2015

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Consumption or mixing of feline ingredients in halal and kosher foods is forbidden, and various diseases such as SARS, anthrax, and hepatitis could be transmitted through feline meats. However, since feline species are abundant across the world without any market price and their meats are consumed in exotic foods, the chances of their adulteration in common meats are very high. Several recent reports appreciated short amplicon-length PCR assays for species authentication in processed foods assuming that shorter targets would be thermodynamically more stable than longer ones under natural decomposition and food processing treatments. However, scientific evidence to prove this hypothesis is rarely documented. For the first time, we developed here a PCR assay targeting only a 69-bp site of feline mitochondrial cytochrome b gene, and its authenticity was confirmed by AluI restriction enzyme followed by its separation and detection on a lab-on-a-chip-based automated electrophoretic system. The exceptional target stability was systematically proven over the previously documented shortest target (108 bp) under extreme autoclaving and microwaving treatments both in pure and mixed matrices. The assay specificity was tested against 14 terrestrial and aquatic species commonly consumed in foods, and no cross-species detection was observed. The limit of detection of the assay was 0.1 pg of feline DNA and 0.01 % (w/w) of feline meats in raw meats and cooked burgers, respectively.

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Species Authentication of Dog, Cat, and Tiger Using Cytochrome β Gene

Cited by 7 publications

References 23 publications

Development of mitochondrial 12S rRNA gene for identification of dog and rat in beef using multiplex PCR

Development of mitochondrial 12S rRNA gene for identification of dog and rat in beef using multiplex PCR

Lab-on-a-chip-based PCR-RFLP assay for the confirmed detection of short-length feline DNA in food

Short Amplicon-Length PCR Assay Targeting Mitochondrial Cytochrome b Gene for the Detection of Feline Meats in Burger Formulation

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