Improved Method for Purification of Bacterial DNA from Bovine Milk for Detection of
            <i>Brucella</i>
            spp. by PCR

Romero, C; López-Goñi, Ignacio

doi:10.1128/aem.65.8.3735-3737.1999

Cited by 67 publications

(30 citation statements)

References 10 publications

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“…Moreover, lysing target cells, especially of Gram‐positive and spore‐forming bacteria, is challenging. In order to overcome the aforementioned drawbacks, several tricks have been developed aimed at i) increasing the numbers of target cells (by introducing a preenrichment step), ii) concentrating and capturing the target cells to isolate them from the matrix, iii) favoring cell lysis through the use of appropriate and, when available, selective lytic enzymes (Romero and Lopez‐Goñi ; Bonaïti and others ; Cremonesi and others ; Fusco and others ,b). When the matrix is known to contain high amounts of milk or dairy matrix‐associated polymerase chain reaction (PCR) inhibitors such as calcium ions and proteinases (McLauchlin and others ; Ramesh and others ; Ercolini and others ; Fusco and others ; Quero and others , b).…”

Section: Culture‐dependent and Culture‐independent Molecular Methods mentioning

confidence: 99%

Culture‐Dependent and Culture‐Independent Nucleic‐Acid‐Based Methods Used in the Microbial Safety Assessment of Milk and Dairy Products

Fusco

Quero

2014

Comp Rev Food Sci Food Safe

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Despite great advances in the diagnostics and better awareness for food safety and security worldwide, significant numbers of foodborne outbreaks have been traced back to the consumption of milk and dairy products contaminated with pathogenic bacteria, such as Listeria monocytogenes, Staphylococcus aureus, Salmonella spp., Campylobacter spp., and pathogenic Escherichia coli. Several culture-dependent and culture-independent nucleic acid-based methods have been proposed to identify, detect, and type milk-and dairyborne pathogenic bacteria. In our review, we will provide an overview on why it is of utmost importance to ascertain the presence of pathogenic microorganisms in milk and milk products; thereafter, we will describe the most commonly used culture-dependent and culture-independent methods, as well as the most attractive ones with regard to their future exploitation, providing the reader with new insights into how and when they can be exploited to ensure the enumeration, and accurate detection at both species and strain level of the most important milk-and dairyborne pathogenic bacteria, even if in a viable but nonculturable state.

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Section: Culture‐dependent and Culture‐independent Molecular Methods mentioning

confidence: 99%

Culture‐Dependent and Culture‐Independent Nucleic‐Acid‐Based Methods Used in the Microbial Safety Assessment of Milk and Dairy Products

Fusco

Quero

2014

Comp Rev Food Sci Food Safe

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“…This occurred despite the fact that primers F4/R2 amplify a region of the 16S rRNA gene present in several copies in the bacterial genome, whereas primers B4/B5 amplify one copy of the gene encoding the 31-kDa B. abortus antigen. Brucella detection limits in cow's milk as reported in the published literature vary greatly, ranging from 2.8 × 10 4 cfu/mL (Rijpens et al, 1996) through 2 × 10 3 cfu/mL (Sreevatsan et al, 2000), to between 5 and 50 cfu/mL (Romero and Lopez-Goni, 1999). In addition, different limits were reported for different species.…”

Section: Discussionmentioning

confidence: 97%

“…We chose 2 pairs of primers: B4/B5, the excellent sensitivity of which was previously reported (Matar et al, 1996;Morata et al, 1998), and F4/R2, which is commonly used for the detection of Brucella spp. in milk (Romero et al, 1995a,b;Romero and Lopez-Goni, 1999). Amplification conditions were based on those used in a previous study, which evaluated the sensitivity of PCR assays using the same primer pairs (Navarro et al, 2002), and on the Taq polymerase manufacturer's instructions.…”

Section: Discussionmentioning

confidence: 99%

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Evaluation of Molecular Methods for the Detection of Brucella Species in Water Buffalo Milk

Marianelli

Martucciello

Tarantino

et al. 2008

Journal of Dairy Science

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Brucellosis is a highly infectious disease affecting both animals and humans. The current standard tools for the diagnosis of this bacterial infection are serological and microbiological. In this study, we evaluated the feasibility of molecular assays as diagnostic tools for the detection of Brucella spp. in water buffalo milk. For this purpose, we first compared different DNA extraction protocols and PCR methods on artificially spiked milk samples. The most sensitive methods were then used to examine milk from serologically positive and negative water buffaloes. Molecular results were compared with serological and bacteriological test results. Milk samples from 53 Brucella seropositive buffaloes (by either rose Bengal or complement fixation test) were positive by ELISA, 37 were positive by culture, 33 were positive by PCR, and 35 were positive by real-time PCR. Of the 37 culture-positive samples, a total of 25 and 26 were positive by PCR and real-time PCR, respectively. Of the 16 culture-negative samples, 8 were positive by PCR and 9 by real-time PCR. Thus, although culture showed greater sensitivity than PCR, some animals found positive by serological methods and PCR tested negative by milk culture. The combined use of bacteriological and molecular tools increased the number of positive samples to 46. In conclusion, these results suggest that the simultaneous application of these 2 direct detection methods (culture and PCR) could be more useful than one test alone for the diagnosis of Brucella spp. in buffalo milk.

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“…paratuberculosis (Johne's disease) , Mycobacterium bovis (Antognoli et al, 2001;Sreevatsan et al, 2000), Brucella spp. (Romero and Lopez-Goñi 1999;Sreevatsan et al, 2000), Coxiella burnetii (Muramatsu et al, 1997), Cryptosporodium spp. (Laberge et al, 1996), and Mycoplasma spp.…”

Section: Pcr/nucleic Acid Tests For the Detection Of Bovine Mastitismentioning

confidence: 99%

Evaluation of a commercial real-time multiplex PCR assay for the detection of bovine mastitis pathogens directly from milk

Lund¹

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Improved Method for Purification of Bacterial DNA from Bovine Milk for Detection of Brucella spp. by PCR

Cited by 67 publications

References 10 publications

Culture‐Dependent and Culture‐Independent Nucleic‐Acid‐Based Methods Used in the Microbial Safety Assessment of Milk and Dairy Products

Culture‐Dependent and Culture‐Independent Nucleic‐Acid‐Based Methods Used in the Microbial Safety Assessment of Milk and Dairy Products

Evaluation of Molecular Methods for the Detection of Brucella Species in Water Buffalo Milk

Evaluation of a commercial real-time multiplex PCR assay for the detection of bovine mastitis pathogens directly from milk

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