2024-03-29T14:26:07Zhttp://digital.csic.es/dspace-oai/requestoai:digital.csic.es:10261/52002021-12-28T16:02:25Zcom_10261_39com_10261_1col_10261_292
Quantitative detection of clostridium tyrobutyricum in milk by real-time PCR
López-Enríquez, Lorena
Rodríguez-Lázaro, David
Hernández, Marta
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.
This work was supported by the Agrarian Experimental Plan of the ITACyL/Junta de Castilla y León and the European Union's Marie-Curie Mobility Program (contract MEIF-CT-2005-0011564). L.L.-E. received a Ph.D. studentship from the Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), D.R.-L. is a fellow of the European Union's Marie-Curie Mobility Program, and M.H. holds a contract from the INIA.
Peer reviewed
2008-06-20T07:50:15Z
2008-06-20T07:50:15Z
2007-06
artículo
http://purl.org/coar/resource_type/c_6501
Applied and Environmental Microbiology 73(11): 3747–3751 (2007)
0099-2240
http://hdl.handle.net/10261/5200
10.1128/AEM.02642-06
17449705
en
http://dx.doi.org/10.1128/AEM.02642-06
open
25095 bytes
application/pdf
American Society for Microbiology