Banner
Current IssueEditorial BoardOnline First Articles
Current IssueEditorial BoardOnline First Articles
Indian Journal of
Animal Research
Print ISSN: 0367-6722
Online ISSN: 0976-0555
NASS Rating
6.40
SJR
0.233
CiteScore
0.606

Chief Editor:
M. R. Saseendranath
Kerala Veterinary and Animal Science University, Mannuthy, Thrissur, INDIA
Frequency:Monthly
Indexing:
Science Citation Index Expanded, BIOSIS Preview, ISI Citation Index, Biological Abstracts, Scopus, ...

Indian Journal of Animal Research, volume 49 issue 4 (august 2015) : 537-541

Rapid identification of goat (Capra hircus) and sheep (Ovis aries) species in raw meat using duplex PCR assay

N. Panwar, G. C. Gahlot, Kritika Gahlot, M. Ashraf, Anoop Singh
1Molecular Genetics Lab, College of Veterinary & Animal Science, Rajasthan University of Veterinary & Animal Sciences, Bikaner 334 003, India.
Cite article:- Panwar N., Gahlot C. G., Gahlot Kritika, Ashraf M., Singh Anoop (2025). Rapid identification of goat (Capra hircus) and sheep (Ovis aries) species in raw meat using duplex PCR assay. Indian Journal of Animal Research. 49(4): 537-541. doi: 10.5958/0976-0555.2015.00077.1.

ABSTRACT

Adulteration of high quality meat with their inferior/cheaper counterparts has become a common practice in the meat industry, which is not detectable by the naked eye or by eating. To circumvent this problem, molecular methods had been developed. The present study was carried out for detection of meat species with the use of cytochrome b gene variability by duplex PCR. Meat samples from goat, sheep and buffalo were utilized for molecular analysis. Genomic DNA was isolated from six samples of each species with some modifications. Mitochondrial cyotchrome b gene was amplified by conventional and duplex PCR using a common forward primer and species-specific reverse primer. PCR amplicons were resolved by agarose gel electrophoresis and for each species produce a characteristic band pattern in conventional and duplex PCR was obtained. The PCR products showed species-specific DNA fragments of 157 and 331 bps from goat and sheep respectively. The duplex PCR could detect upto 10 % of DNA of meat species. Thus, the duplex PCR was found to be a simple, reliable, sensitive and highly specific test for simultaneous detection of meat species.

REFERENCES

  1. Ausubel, F. M., Brent, R., Kingston, R. E., Moore, D. D., Seidman, J. G., Smith, J. A. and Strehl, K. (1987). Current protocols in molecular biology. Green Publishing Associates and Wiley- Interscience, Newyork, USA, pp 2.2.1.
  2. Che Man, Y.B., Mustafa, S., Mokhtar, N.F.K., Nordin, R. and Sazili, A.Q. (2012). Porcine-specific polymerase chain reaction assay based on mitochondrial d-loop gene for identification of pork in raw meat. Int. J. Food Properties. 15:134-144.
  3. Chikuni K., Tabata T., Kosugiyama M., Monma M., Saito M., (1994). Polymerase chain reaction assay for detection of sheep and goat meats. Meat Science. 37:337-345.
  4. Deepak Kumar, Singh S. P., Nagappa S. Karabasanavar, Rashmi Singh, Umapathi V. (2012). Authentication of beef, carabeef, chevon, mutton and pork by a PCR-RFLP assay of mitochondrial cytb gene. Journal of Food Science and Technology 49: 542-48.
  5. Fei S., Okayama T., Yamanoue M., Nishikawa I., Mannen H., Tsuji S., (1996). Species identification of meats and meat products by PCR. Animal Science and Technology (Jpn), 67: 900-905.
  6. Girish PS, Anjaneyulu AS, Viswas KN, Shivakumar BM, Anand M, Patel M, Sharma B. (2005). Meat species identification by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) of mitochondrial 12S rRNA gene. Meat Science 70:107-12.
  7. Henegariu O, Heerema NA, Dlouhy SR, Vance GV, Vogt PH (1997). Multiplex PCR: Critical Parameters and Step-by-Step Protocol. BioTechniques, 23: 504-511.
  8. Hopwood, A. J., Fairbrother, K. S., Lockley, A. K. and Bardsley, R.G. (1999). An actin gene related polymerase chain reaction test for identification of chicken in meat mixtures. Meat Sci. 53: 227- 231.
  9. Hsieh HM, Chiang HL, Tsai LC, Lai SY, Huang NE, Linacre A (2001). Cytochrome b gene for species identification of the conservation animals. Forensic Sci. Int. 122: 7-18.
  10. Kesmen Z, Gulluce A, Sahin F, Yetim H (2009). Identification of meat species by TaqMan-based real-time PCR assay. Meat Sci. 82: 444-449.
  11. Kumar D, Singh SP, Singh R, Karabasanavar NS (2011). A highly specific PCR assay for identification of goat (Capra hircus) meat. Small Rumin Res. 97:76–78
  12. Lockley A.K., Bardsley R.G., (2000). DNA – based methods for food authentication. Trends Food Sci. Technol. 11: 67-77.
  13. Lopez-Andreo M, Lugo L, Garrido-Pertierra A, Isabel Prieto M, Puyet A (2005). Identiûcation and quantitation of species in complex DNA mixtures by real-time polymerase chain reaction. Anal. Biochem. 339: 73-82.
  14. Maede D (2006) A strategy for molecular species detection in meat and meat products by PCR–RFLP and DNA sequencing using mitochondrial and chromosomal genetic sequences. European Food Research and Technology 224:209–217
  15. Malisa, A., Gwakisa, P., Balthazary, S., Wasser, S.K. and Mutayaba, B. (2006) The potential of mitochondrial DNA markers and PCR-RFLP for domestic and wild species identification. African Journal of Biotech. 5:1588-1593.
  16. Mane, B.G., Mendiratta, S.K. and Tiwari, A.K. (2009). Polymerase chain reaction assay for identification of chicken in meat and meat products. Food Chemistry. 116:806–810.
  17. Matasunaga, T., K. Chikuni, R. Tanabe, S. Muroya, K. Shaibata, J. Yamaa, and Y. Shinmura, (1999). A quick and simple method for the identification of meat species and meat products by PCR assay. Meat Science 51:143-148.
  18. Meyer, R., Candrian, U. and Luthy, J. (1994). Detection of pork in heated meat products by polymerase chain reaction. JAOAC Inter. 77:617-622.
  19. Meyer R., Hofelein C., Luthy J., Candrian U., (1995). Polymerase chain reaction-restriction fragment length polymorphism analysis: a simple method for species identification food. Journal Association of Official Analytical Chemists. 78:1542-1551.
  20. Partis L, Croan D, Guo Z, Clark R, Coldham T, Murby J (2000). Evaluation of a DNA fingerprinting method for determining the species origin of meats. Meat Sci. 54: 369-376.
  21. Satish S, Ashwani kumar, Y Singh Y. Minakshi and Gaya Prasad (2011). Identification and differentiation of raw meats of sheep, goat, cattle, pig and poultry by multiplex PCR. Indian Journal of Animal Sciences 81: 588–591.
  22. Spychaj A., Mozdziak P.E., Pospiech E., (2009). PCR methods in meat species identification as a tool for the verification of regional and traditional meat products. Acta Sci.Pol., Technol. Aliment. 8: 5-20.
  23. Sun YL and Lin CS (2003). Establishment and application of a fluorescent polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method for identifying porcine, caprine, and bovine meats. J Agric Food Chem 51:1771-6.
  24. Zarringhabaie, G.E., Pirany, N. and Javanmard, A. (2011). Molecular traceability of the species origin of meats using multiplex PCR. African Journal of Biotechnology 10:15461-16465.
  25. Zhang G.L., Zheng M.G., Zhou Z.J., Ouyang H.S., LU Q., (1999). Establishment and application of a polymerase chain reaction for the identification of beef. Meat Science 51, 233-236.
Reviewed By

Download Article
In this Article
    Contact us
    Follow us

    Editorial Board

    View all (0)