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Title

Quantitative detection of clostridium tyrobutyricum in milk by real-time PCR

AuthorsLópez-Enríquez, Lorena; Rodríguez-Lázaro, David; Hernández, Marta
Issue DateJun-2007
PublisherAmerican Society for Microbiology
CitationApplied and Environmental Microbiology 73(11): 3747–3751 (2007)
AbstractWe 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 (R2 > 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.
Publisher version (URL)http://dx.doi.org/10.1128/AEM.02642-06
URIhttp://hdl.handle.net/10261/5200
DOI10.1128/AEM.02642-06
ISSN0099-2240
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