Real-time TaqMan PCR for quantitative detection of cows’ milk in ewes’ milk mixtures

Real-time TaqMan PCR for quantitative detection of cows’ milk in ewes’ milk mixtures A real-time PCR based on the amplification of a fragment of the mitochondrial 12S ribosomal RNA gene was developed and evaluated for the detection and quantification of cows’ milk in raw and heat-treated cow/sheep milk mixtures. The method combines the use of cow-specific primers that amplify a 252 bp fragment from cow DNA, and mammalian-specific primers amplifying a 428 bp fragment from mammalian species DNA, which is used as an endogenous control. The method measures PCR product accumulation through a 6-carboxyfluorescein-labeled fluorogenic probe (TaqMan). A comparison of the cycle number at which mammalian and cow-specific PCR products were first detected, in combination with the use of reference standards of known bovine content, allowed the determination of the percentage of cows’ milk in mixtures. Experimental raw and heat-treated binary mixtures were analyzed, demonstrating the specificity and sensitivity of the assay for detection and quantification of cows’ milk in the range 0.5–10%. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png International Dairy Journal Elsevier

Real-time TaqMan PCR for quantitative detection of cows’ milk in ewes’ milk mixtures

Loading next page...
 
/lp/elsevier/real-time-taqman-pcr-for-quantitative-detection-of-cows-milk-in-ewes-fGayE7FW7S
Publisher
Elsevier
Copyright
Copyright © 2006 Elsevier Ltd
ISSN
0958-6946
eISSN
1879-0143
D.O.I.
10.1016/j.idairyj.2006.09.005
Publisher site
See Article on Publisher Site

Abstract

A real-time PCR based on the amplification of a fragment of the mitochondrial 12S ribosomal RNA gene was developed and evaluated for the detection and quantification of cows’ milk in raw and heat-treated cow/sheep milk mixtures. The method combines the use of cow-specific primers that amplify a 252 bp fragment from cow DNA, and mammalian-specific primers amplifying a 428 bp fragment from mammalian species DNA, which is used as an endogenous control. The method measures PCR product accumulation through a 6-carboxyfluorescein-labeled fluorogenic probe (TaqMan). A comparison of the cycle number at which mammalian and cow-specific PCR products were first detected, in combination with the use of reference standards of known bovine content, allowed the determination of the percentage of cows’ milk in mixtures. Experimental raw and heat-treated binary mixtures were analyzed, demonstrating the specificity and sensitivity of the assay for detection and quantification of cows’ milk in the range 0.5–10%.

Journal

International Dairy JournalElsevier

Published: Jul 1, 2007

References

  • Application of immunological methods for the detection of species adulteration in dairy products
    Hurley, I.P.; Ireland, H.E.; Coleman, R.C.; Williams, J.H.H.
  • PCR detection of cow's milk in water buffalo milk and mozzarella cheese
    López-Calleja, I.; González, I.; Fajardo, V.; Rodríguez, M.A.; Hernández, P.E.; García, T.
  • Milk species identification in cheese varieties using electrophoretic, chromatographic and PCR techniques
    Mayer, H.K.
  • Recent developments in antibody-based analytical methods for the differentiation of milk from different species
    Moatsou, G.; Anifantakis, E.
  • Real-time PCR for quantitative meat species testing
    Sawyer, J.; Wood, C.; Shanahan, D.; Gout, S.; McDowell, D.

You’re reading a free preview. Subscribe to read the entire article.


DeepDyve is your
personal research library

It’s your single place to instantly
discover and read the research
that matters to you.

Enjoy affordable access to
over 18 million articles from more than
15,000 peer-reviewed journals.

All for just $49/month

Explore the DeepDyve Library

Search

Query the DeepDyve database, plus search all of PubMed and Google Scholar seamlessly

Organize

Save any article or search result from DeepDyve, PubMed, and Google Scholar... all in one place.

Access

Get unlimited, online access to over 18 million full-text articles from more than 15,000 scientific journals.

Your journals are on DeepDyve

Read from thousands of the leading scholarly journals from SpringerNature, Elsevier, Wiley-Blackwell, Oxford University Press and more.

All the latest content is available, no embargo periods.

See the journals in your area

DeepDyve

Freelancer

DeepDyve

Pro

Price

FREE

$49/month
$360/year

Save searches from
Google Scholar,
PubMed

Create folders to
organize your research

Export folders, citations

Read DeepDyve articles

Abstract access only

Unlimited access to over
18 million full-text articles

Print

20 pages / month

PDF Discount

20% off