Polymerase Chain Reaction–Restriction Fragment Length Polymorphism of Mitochondrial 12S rRNA Gene: A Simple Method for Identification of Poultry Meat Species

Girish, P. S.; Anjaneyulu, A. S. R.; Viswas, K.N.; Santhosh, F. H.; Bhilegaonkar, K. N.; Agarwal, R. K.; Kondaiah, N.; Nagappa, K.

doi:10.1007/s11259-006-3390-5

Cited by 43 publications

(25 citation statements)

References 11 publications

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“…Earlier workers have also exploited mitochondrial 12S rRNA gene (Prakash et al 2000;Girish et al 2004;Rastogi et al 2004) and mitochondrial cytochrome b gene (Meyer et al 1995;Unseld et al 1995;Hsieh et al 2005) for meat species identification, and it has also been reported that the PCR amplification of mitochondrial 16S rRNA gene has been used for identification of snail species (Abdulmawjood and Buelte, 2002). Earlier also reported that the PCR amplification followed by DNA sequencing can be the best way to identify the species origin of meat from all meat animals (Girish et al 2007). …”

Section: Resultsmentioning

confidence: 99%

“…Identification of buffalo, emu and crocodile species was done by PCR amplification and sequencing of cytochrome b gene (Forrest and Carnegie 1994). Recently, Girish et al (2007) employed partial sequence analysis of 402 bp PCR amplified fragments of cytochrome b gene for identification of animal species in fresh and processed samples of unknown origin. These workers identified the species in unknown samples by the sequence analysis of PCR amplified DNA fragments.…”

Section: Introductionmentioning

confidence: 99%

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Sequence analysis of mitochondrial 16S rRNA gene to identify meat species

Mane

Mendiratta

Tiwari

et al. 2013

Journal of Applied Animal Research

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In the present study, mitochondrial 16S rRNA gene sequence analysis was used for identification of Cattle (Bos taurus), buffalo (Bubalus bubalis), sheep (Ovis aries), goat (Capra hircus) and pig (Sus scrofa) species in fresh and processed meat. The DNA was extracted from fresh and processed meat including autoclaved meat and meat emulsion followed by polymerase chain reaction (PCR) amplification of about 497 bp DNA fragments of mitochondrial 16S rRNA gene. Then the amplified PCR fragments were sequenced and analysed to differentiate the species. No adverse effect of ingredients and processing conditions was observed on PCR amplification of DNA extracted from heat-treated meat and meat emulsion. The closely related species such as cattle and buffalo, sheep and goat were differentiated from each other by sequence analysis. Thus, PCR amplification and sequence analysis of mitochondrial 16S rRNA gene can be used as a tool for authentic identification of meat species.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Sequence analysis of mitochondrial 16S rRNA gene to identify meat species

Mane

Mendiratta

Tiwari

et al. 2013

Journal of Applied Animal Research

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“…Compared to other meat species, there are fewer poultry-specific assays for food authentication and quality control (Stamoulis et al 2010). Most poultry DNA-based assays utilize avian mitochondrial genes and do not have a nuclear DNA quantification component (Dalmasso et al 2004;Girish et al 2007;Stamoulis et al 2010;Kocher et al 1989;Herman 2004;Haunshi et al 2009;Girish et al 2011). The integration of species determination and nuclear DNA quantification into a single assay will streamline downstream genotyping analysis, and improve the quality of the DNA profiles while conserving reagents (Evans et al 2007;Lindquist et al 2011;Kanthaswamy et al 2012).…”

Section: Introductionmentioning

confidence: 99%

“…Of the few poultry assays developed, most rely on mitochondrial DNA (mtDNA) genes as markers for detection (Girish et al 2007;Haunshi et al 2009;Herman 2001;Soares et al 2010). Since mtDNA occurs in high copy numbers in each cell and is able to withstand degradation and environmental challenges, it is well suited for use in food authentication assays.…”

Section: Introductionmentioning

confidence: 99%

A nuclear DNA-based species determination and DNA quantification assay for common poultry species

Satkoski

Premasuthan

et al. 2012

J Food Sci Technol

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DNA testing for food authentication and quality control requires sensitive species-specific quantification of nuclear DNA from complex and unknown biological sources. We have developed a multiplex assay based on TaqMan® realtime quantitative PCR (qPCR) for species-specific detection and quantification of chicken (Gallus gallus), duck (Anas platyrhynchos), and turkey (Meleagris gallopavo) nuclear DNA. The multiplex assay is able to accurately detect very low quantities of species-specific DNA from single or multispecies sample mixtures; its minimum effective quantification range is 5 to 50 pg of starting DNA material. In addition to its use in food fraudulence cases, we have validated the assay using simulated forensic sample conditions to demonstrate its utility in forensic investigations. Despite treatment with potent inhibitors such as hematin and humic acid, and degradation of template DNA by DNase, the assay was still able to robustly detect and quantify DNA from each of the three poultry species in mixed samples. The efficient species determination and accurate DNA quantification will help reduce fraudulent food labeling and facilitate downstream DNA analysis for genetic identification and traceability. ). The integration of species determination and nuclear DNA quantification into a single assay will streamline downstream genotyping analysis, and improve the quality of the DNA profiles while conserving reagents (Evans et al. 2007;Lindquist et al. 2011;Kanthaswamy et al. 2012). Many of the DNA-based food authentication assays are multiplexed end-point PCR assays (Dalmasso et al. 2004;Zha et al. 2010;Ghovvati et al. 2009;Rodriguez et al. 2003). A majority of these protocols require subsequent steps such as restriction enzyme digestion or gel electrophoresis for species identification after PCR. Endpoint PCR assays with a quantification component have been developed, but amplicon quantities are typically determined using imaging software analysis of fluorescence intensities of electrophoresis gel bands (Soares et al. 2010).Like end-point PCR, real-time quantitative PCR (qPCR) technology allows multiplexing, and is amenable to detecting multiple DNA targets that allow the simultaneous identification of DNA originating from multiple species. Moreover, the real-time DNA quantification feature of qPCR technology facilitates the efficient and accurate quantification of template DNA. While simplifying data collection and analysis, qPCR's ability to amplify, identify, and quantify in a single step effectively minimizes turnaround time and exposure to contamination and errors ).Electronic supplementary material The online version of this article

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“…The DNA molecule is more stable and the composition of DNA is same in any cell of the individual. Hence, DNA based techniques namely Polymerase Chain Reaction (PCR) and its variants: Restriction Fragment Length Polymorphism (RFLP) (Girish et al 2005); Random Amplified Polymorphic DNA (RAPD) finger printing (Saez et al 2004); DNA Hybridization (Buntjer et al 1999), PCR Sequencing (Girish et al 2007); Arbitrarily Primed-PCR (AP-PCR) (Desmarais et al 1998) and PCR-Single Strand Conformation Polymorphisms (SSCP) (Tejedor et al 2006) and species specific markers (Haunshi et al 2009) have been employed for the identification of various animal species in the recent past. The PCR-based techniques provide high level of specificity, sensitivity, accuracy and precision than the techniques hitherto being used for the purpose of animal species identification.…”

Section: Introductionmentioning

confidence: 99%

A rapid method for authentication of Buffalo (Bubalus bubalis) meat by Alkaline Lysis method of DNA extraction and species specific polymerase chain reaction

et al. 2011

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Buffalo (Bubalus bubalis) meat is a major item of export from India but export of beef i.e. meat from cattle (Bos indicus) is prohibited. Also, adulteration of buffalo meat with that of beef (meat from cattle) is a common fraudulent practice because of prohibition on cow slaughter in most states of India. Food analysts require precise identification techniques to implement such regulations. In the present study, a method of DNA extraction by Alkaline lysis from meat samples and speciation of buffalo meat using species specific Polymerase Chain Reaction (PCR) has been reported. Alkaline lysis technique is a rapid method which involves triturating meat with four volumes of 0.2N NaOH, dilution of resultant liquid extract with eight volumes of 0.2N NaOH, heating the mix 75°C for 20 min followed by neutralization with eight volumes of 0.04N Tris HCl. Entire procedure of DNA extraction takes less than 30 min and it is economical as it involves less expensive chemicals. Method was successfully applied in animal byproducts also viz., liver, heart and kidney. For authentication of buffalo meat, pair of primers was designed based on mitochondrial D loop gene nucleotide sequence. PCR amplification using the designed primers gave amplicon of size 482 bp in buffalo and no amplification was detected in closely related species viz., cattle, sheep and goat meat samples. Results of the assay were highly repetitive and reliable. An export sample referred by export regulation authorities was also analyzed by using the Alkaline lysis method of DNA extraction and species specific PCR which enabled authentication of meat within 5 h.

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Polymerase Chain Reaction–Restriction Fragment Length Polymorphism of Mitochondrial 12S rRNA Gene: A Simple Method for Identification of Poultry Meat Species

Cited by 43 publications

References 11 publications

Sequence analysis of mitochondrial 16S rRNA gene to identify meat species

Sequence analysis of mitochondrial 16S rRNA gene to identify meat species

A nuclear DNA-based species determination and DNA quantification assay for common poultry species

A rapid method for authentication of Buffalo (Bubalus bubalis) meat by Alkaline Lysis method of DNA extraction and species specific polymerase chain reaction

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