title: Development of a real-time PCR assay for direct detection and quantification of Bacillus sporothermodurans in ultra-high temperature milk.
abstract: In our study, a new and highly sensitive real-time PCR Taqman assay was developed for the direct and specific detection of Bacillus sporothermodurans in UHT milk. The target region was selected based on the 16S rRNA gene profiles of 11 B. sporothermodurans from GenBank. A standard curve was created using a reference strain of B. sporothermodurans, DSM 10599. A low limit of detection for B. sporothermodurans in UHT milk (10 cfu/mL) was obtained. Furthermore, a total of 110 UHT milk samples from several supermarkets were directly assessed to detect and quantify B. sporothermodurans using the real-time PCR Taqman developed. The B. sporothermodurans counts obtained were highly correlated with the microbial plate counts in the UHT milk samples. This is the first time that B. sporothermodurans has been quantified directly from UHT milk. This technique could be applicable as a routine tool for preventing the growth of these bacteria by allowing for the rapid screening of raw milk samples in dairy plants. As expected, the probability of bacterial growth in UHT milk packages increased with the B. sporothermodurans counts in the raw milk.
authors: Department of Analytical Chemistry, Nutrition and Food Science, Faculty of Veterinary, University of Santiago de Compostela, Lugo 27002, Spain; Dairy Department, Faculty of Agriculture, Suez Canal University, Ismailia 41522, Egypt.
authors: M
authors: M
authors: Abouelnaga
authors: Department of Analytical Chemistry, Nutrition and Food Science, Faculty of Veterinary, University of Santiago de Compostela, Lugo 27002, Spain.
authors: A
authors: A
authors: Lamas
authors: Department of Analytical Chemistry, Nutrition and Food Science, Faculty of Veterinary, University of Santiago de Compostela, Lugo 27002, Spain.
authors: J M
authors: JM
authors: Miranda
authors: Dairy Department, Faculty of Agriculture, Suez Canal University, Ismailia 41522, Egypt.
authors: M
authors: M
authors: Osman
authors: Department of Analytical Chemistry, Nutrition and Food Science, Faculty of Veterinary, University of Santiago de Compostela, Lugo 27002, Spain.
authors: A
authors: A
authors: Cepeda
authors: Department of Analytical Chemistry, Nutrition and Food Science, Faculty of Veterinary, University of Santiago de Compostela, Lugo 27002, Spain. Electronic address: carlos.franco@usc.es.
authors: C M
authors: CM
authors: Franco
mesh_terms: D000818:Animals; D001407:Bacillus / Q000145:classification*; D008892:Milk / Q000382:microbiology*; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D013696:Temperature
chemical_list: D012336:RNA, Ribosomal, 16S
keywords: Bacillus sporothermodurans; real-time PCR Taqman; ultra-high temperature milk