Quantitative Detection of Clostridium tyrobutyricum in Milk by Real-Time PCR |
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| Authors: | Lorena L pez-Enrí quez, David Rodrí guez-L zaro, Marta Hern ndez |
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| Affiliation: | Lorena López-Enríquez, David Rodríguez-Lázaro, and Marta Hernández |
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| 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 (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. |
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