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Appl Environ Microbiol. 2007 Jun;73(11):3747-51. Epub 2007 Apr 20.

Quantitative detection of Clostridium tyrobutyricum in milk by real-time PCR.

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1
Instituto Tecnológico Agrario de Castilla y León, Carretera de Burgos, km. 119, 47071 Valladolid, Spain.

Abstract

We developed a real-time PCR assay for the quantitative detection of Clostridium tyrobutyricum, which has been identified as the major causal agent of late blowing in cheese. The assay was 100% specific, with an analytical sensitivity of 1 genome equivalent in 40% of the reactions. The quantification was linear (R(2) > 0.9995) over a 5-log dynamic range, down to 10 genome equivalents, with a PCR efficiency of >0.946. With optimized detergent treatment and enzymatic pretreatment of the sample before centrifugation and nucleic acid extraction, the assay counted down to 300 C. tyrobutyricum spores, with a relative accuracy of 82.98 to 107.68, and detected as few as 25 spores in 25 ml of artificially contaminated raw or ultrahigh-temperature-treated whole milk.

PMID:
17449705
PMCID:
PMC1932699
DOI:
10.1128/AEM.02642-06
[Indexed for MEDLINE]
Free PMC Article
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