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Meat Species Identification: Amplification Refractory Mutation System-Polymerase Chain Reaction–Based Assay

  • Muhammad WaqasEmail author
  • Zahid Hussain
  • Awais Ihsan
Article
  • 19 Downloads

Abstract

True verification of meat species is critical for religious, economical, legal, and/or public health concerns. Current methods available for verification of meat species are very time-consuming and/or costly. So considering the necessity, a novel polymerase chain reaction assay (PCR)–based ARMS (amplification refractory mutation system) type identification assay for detection of 6 most common mammalian species—Equus asinus (donkey), Equus caballus (horse), Bos Taurus (cow), Bubalus bubalis (buffalo), Capra hircus (goat), and Canis lupus familiaris (dog)—was described in present study by designing species-specific forward primers against variable regions and a single universal reverse primer against a very conserved region of nuclear beta actin (ACTB) gene by investigation of areas of homology and variation. As compared with conventional designing of primers, ARMS type primer designing is a better alternative as it allows more room for choice and offers differentiation of closely related species by exploiting just a single nucleotide base difference. PCR bands of 128, 229, 273, 362, 710, and 796 bp were generated on electrophoretic gel for buffalo, donkey, cow, horse, dog, and goat meat species respectively. Besides singleplexing, duplex (multiplex) PCR for donkey and horse, donkey and goat, and donkey and buffalo were also performed which successfully generated corresponding bands. The method is a simple and straightforward setup; results can be interpreted easily in a short time and do not need validation by sequencing. Results of the present study clearly demonstrate that the method can be used as an identification tool for differentiation between cow, buffalo, goat, donkey, horse, and dog species.

Keywords

ACTB gene (beta actin) ARMS (amplification refractory mutation system) PCR (polymerase chain reaction) Meat adulteration Meat authenticity Species identification 

Notes

Acknowledgments

The authors would like to thank Higher Education Commission (HEC) Pakistan, GCU Lahore Pakistan and COMSATS Institute of Information Technology Sahiwal for financial and experimental support. We would also like to express special thanks to Dr. Sumaira Kousar (GCWU, Faisalabad) and Dr. Waqar Anjum (UVAS, Lahore, Pakistan) for their cooperation and support in providing samples for the research.

Compliance with Ethical Standards

Conflict of Interest

Muhammad Waqas declares that there is no conflict of interest. Zahid Hussain declares that there is no conflict of interest. Awais Ihsan declares that there is no conflict of interest.

Ethical Approval

All applicable international, national, and/or institutional guidelines for the care and use of animals were followed.

Informed Consent

Not applicable.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Institute of Industrial BiotechnologyGovernment College UniversityLahorePakistan
  2. 2.Department of BiosciencesCOMSATS Institute of Information TechnologySahiwalPakistan

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