PCR based assay for the detection of cow milk adulteration in buffalo milk 
 

DOI: 10.18805/ijar.11331    | Article Id: B-3232 | Page : 383-387
Citation :- PCR based assay for the detection of cow milk adulteration in buffalo milk .Indian Journal Of Animal Research.2018.(52):383-387

Tanmay Hazra, Vivek Sharma, Rekha Sharma and Sumit Arora 

tanmayhazra08@gmail.com
Address :

Dairy Chemistry Division, ICAR-National Dairy Research Institute Karnal-132001, Haryana, India.

Submitted Date : 27-04-2016
Accepted Date : 14-06-2016

Abstract

It is very common in India that with low priced cow milk adulterated high priced buffalo milk. Various techniques are available in market to identify the origin of milk but all these techniques have their own limitations. Thus, DNA based technologies are preferred now a days to identify the origin of food products from animal sources due to high sensitivity and specificity. Currently a PCR based method was developed to identify cow milk in raw buffalo milk. DNA was isolated from milk by DNeasy Mericon food kit (Quiagen,USA) which resulted in DNA of requisite quality for downstream applications. A bovine specific primer targeting D-loop (displacement) of mt- DNA (mitochondrial) was selected and standardized to amplify cow DNA. Specificity of primer was tested across the species in the genomic DNA isolated from both milk and blood. The protocol can be sensitive to detect upto 5% level of cow milk in the buffalo milk. Designed protocol was efficient, robust and sensitive and could be used as a platform test in routine quality analysis laboratory. Hence, it could be a great alternative for other protein based methods to identify cow milk in buffalo milk.

Keywords

Adulteration Buffalo milk Cow milk Mitochondrial DNA Silica column PCR.

References

  1. Calleja,I.L., Alonso,G.I.,Fajardo, V., Rodriguez,M.A .,HernAndez,P.A., Garcia,T. and Martýin,R.(2005) PCR detection of cows’ milk in water buffalo milk and mozzarella cheese. Int. Dairy J., 15:1122-1129.
  2. Cheng,Y., Chen,S. and Weng,C.(2006). Investigation of Goats’ Milk Adulteration with Cows’ Milk by PCR. Asian-Aust. J. Anim. Sci,19 : 1503 – 1507.
  3. De, S., Brahma, B., Polley, S. ,Mukherjee, A. ,Banerjee, D. and Gohaina, M. (2011) Simplex and duplex PCR assays for species specific identification of cattle and buffalo milk and cheese. Food Cont., 22:690–696.
  4. Girish,P.S., Haunshi,S., Vaithiyanathan,S., Rajitha,R., Ramakrishna,R.(2013). A rapid method for authentication of Buffalo (Bubalus bubalis) meat by Alkaline Lysis method of DNA extraction and species specific polymerase chain reaction. J Food Sci Technol, 50: 141–146.
  5. Golinelli , L. P., Carvalho , A. C., Casaes , R. S., Lopes , C. S. C., Deliza , R. , Paschoalin , V. M. F. and Silva , J. T. (2014). Sensory analysis and species-specific PCR detect bovine milk adulteration of frescal (fresh) goat cheese. J. Dairy Sci., 95 : 2749–2752
  6. Kocher, T. D., Thomas, W.K., Meyer. A., Edwards, S.V., Pabo, S., Villablanca, F.X. and Wilson, A. C. (1989). Dynamics of mitochondrial DNA evolution in mammals: amplification & sequencing with conserved primers. Proc. Natl. Acad. Sci. 86: 6196- 6200
  7. Kotowicz,M., Adamczyk,E. and Bania,J .(2007). Application of a duplex-pcr for detection of cows’ milk in goats’ milk. Ann. Agric. Environ. Med., 14: 215-218.
  8. Lipkin, E., Shalom, A., Khatib, H., Soller, M. and Friedmann, A. (1993) Milk as a source of deoxyribonucleic acid and as a substrate for the polymerase chain reaction. J. of Dairy Sci., 76: 2025–2032.
  9. Mane,B.G., Mendiratta, S.K. and Tiwari, A.K. (2012). Beef specific polymerase chain reaction assay for authentication of meat and meat products. Food Cont., 2:246-249.
  10. Maudet, C and Taberlet, P (2001). Detection of cows’ milk in goats’ cheeses inferred from mitochondrial DNA polymorphism. J. of Dairy Res., 68: 229-235.
  11. Nooratiny, I., Sahilah, A.M., Alif Alfie, A. R., Mohd and Farouk, M. Y.(2013) .DNA extraction from ghee and beef species identification using polymerase chain reaction (PCR) assay. Int. Food Res J., 20: 2959-2961.
  12. Pirondini,A.,Bonas,U., Maestri,M., Giovanna,V., Marta,M. and Nelson,M.(2010). Yield and amplificability of different DNA extraction procedures for traceability in the dairy food chain. Food Cont., 21: 663–668.
  13. Rea, S., Chikuni, K., Branciari, R., Sukasi Sangamayya, R., Ranucci, D. and Avellini, P. (2001). Use of duplex polymerase chain reaction (duplex PCR) technique to identify bovine and water buffalo milk used in making mozzarella cheese. J. of Dairy Res., 68: 689–698
  14. Woolfe, M. and Primrose, S. (2004)Food forensics: Using DNA technology to combat misdescription and fraud. Trends in Bioteh., 22: 222–226.
  15. Zachar,P., Michal,S., Radovan,K.,Jarosla,N., Miroslava,N. and Dana,M.(2011)Analytical methods for the species identification of milk and milk products. Mljekarstvo., 61: 199-207.
  16. Zhang, C., Fowler, R.M., Scott, N., Lawson, G. and Slater,A.(2007). A TaqMan real-time PCR system for the identification and quantification of bovine DNA in meats, milks and cheeses. Food Cont., 18: 149–1158.
     

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