LAMP assay for rapid diagnosis of cow DNA in goat milk and meat samples

Document Type : Short paper

Authors

1 Molecular Genetics Laboratory, ICAR-Central Institute for Research on Cattle, Meerut-250 001, Uttar Pradesh, India

2 MSc in Biotechnology, Molecular Genetics Laboratory, ICAR-Central Institute for Research on Cattle, Meerut-250 001, Uttar Pradesh, India

Abstract

Animal species detection is one of the crucial steps for consumer’s food analysis. In the present study we developed an in-house built loop-mediated isothermal amplification (LAMP) assay for rapid detection of adulterated cow DNA in goat milk/meat samples. The cow milk/tissue DNA in goat milk/meat samples were identified in the developed LAMP assay by either naked eye visualizing with SYBR Green I dyes or by detecting the typical ladder pattern on gel electrophoresis. This test can detect up to minimum 5% level of cow components admixed in goat milk/meat samples and can be completed within 1 h 40 min starting from DNA extraction from milk/meat samples and can be performed in a water bath. Developed LAMP methodology is simple; rapid and sensitive techniques that can detect adulterant like cow components in goat milk/meat are more accurate than other existing DNA based technologies.

Keywords

References

Abdel-Rahman, SM and Ahmed, MMM (2007). Rapid and sensitive identification of buffalo’s, cattle’s and sheep’s milk using species-specific PCR and PCR-RFLP tech-niques. Food Control. 18: 1246-1249.
Abdulmawjood, A; Grabowski, N; Fohler, S; Kittler, S; Nagengast, H and Klein, G (2014). Development of loop-mediated isothermal amplification (LAMP) assay for rapid and sensitive identification of ostrich meat. PLoS One. 25, 9: e100717.
Bitri, L; Rolland, MP and Besancon, P (1993). Immuno-logical detection of bovine caseinomacropeptide in ovine and caprine dairy products. Milchwissenschaft. 48: 367-371.
Calvano, CD; De, CC; Monopoli, A and Zambonin, CG (2012). Detection of sheep and goat milk adulterations by direct MALDI-TOF MS analysis of milk tryptic digests. J. Mass Spectrom. 47: 1141-1149.
Deb, R; Sengar, G S; Singh, U; Kumar, S; Alyethodi, RR; Alex, R; Raja, T V; Das,AK; Prakash, B (2016). Application of a loop-mediated isothermal amplification assay for rapid detection of cow components adulterated in buffalo milk/meat. Molecular Biotechnology.doi:10.1007/ s12033-016-9984-4.
Khanzadi, S; Jamshidi, A; Razmyar, J and Mohsenzadeh, M (2013). PCR-based detection of cow and goat milk in sheep milk and dairy‎ products marketed in Mashhad city of Iran. Iranian J. Vet. Med., 7: 257-262.
Lee, CC; Chang, HS and Sheen, HS (2004). A quick novel method to detect the adulteration of cow milk in goat milk. Asian-Aust. J. Anim. Sci., 17: 420-422.
Lopez-Calleja, I; Gonzalez, I; Fajardo, V; Rodriguez, MA; Hernandez, PE; Garcia, T and Martin, R (2004). Rapid detection of cows’ milk in sheeps’ and goats’ milk by a species-specific polymerase chain reaction technique. J. Dairy Sci., 87: 2839-2845.
Notomi, T; Okayama, H; Masubuchi, H; Yonekawa, T; Watanabe, K; Amino, N and Hase, T (2000). Loop-mediated isothermal amplification of DNA. Nucl. Acids Res., 28: e63-e63.
Pawar, SS; Meshram, CD; Singh, NK; Sonwane, AA; Saini, MR; Muglikar, DM; Mishra, BP and Gupta, PK (2014). Rapid detection of bovine herpesvirus 1 in bovine semen by loop-mediated isothermal amplification (LAMP) assay. Arch Virol., 159: 641-648. doi: 10.1007/s00705-013-1869-2.
Volk, H; Klančnik, A; Toplak, N; Kovač, M and Jeršek, B (2014). Real-time PCR detection of fraudulent addition of bovine milk to caprine and ovine milk. J. Hyg. Eng. Design. 7: 107-111.
Wang, D (2015). Novel primers for increased specificity and sensitivity for the detection of Staphylococcus aureus by real-time LAMP. J. Food. doi: 10.1080/19476337.2015. 1048530.
Zhang, CL; Fowler, MR; Scott, NW; 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 Control. 18: 1149-1158.
 

Files

Share

How to cite

Statistics