Direct detection of Listeria monocytogenes from milk by magnetic based DNA isolation and PCR

Amagliani, Giulia; Brandi, Giorgio; Omiccioli, Enrica; Casiere, Annarita; Bruce, Ian J.; Magnani, Mauro

doi:10.1016/j.fm.2003.10.008

Cited by 71 publications

(49 citation statements)

References 24 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The hlyA gene region is known to be specific for L. monocytogenesagainst other Listeria spp. which were included in Amagliani et al 2004and Choi and Hong 2003. Also, Meloni et al (2014 reported that the detection of L. monocytogenes for tackling the trouble in the manufacturing process was accomplished in fermented sausages using 10 genes including hlyA.…”

Section: Specificity Of Primersmentioning

confidence: 99%

A Rapid and Cost‐Efficient Technique for Simultaneous/Duplex Detection of Listeria Monocytogenes and Escherichia Coli O157:H7 Using Real Time PCR

Kaynak

Şakalar

2016

Journal of Food Safety

View full text Add to dashboard Cite

The increasing industrial interest in easy, economical and reliable methods has led to the development and application of DNA-based methods for the detection of microbial pathogens in food. In the present article, we describe the development of a cost-efficient EvaGreen-based real-time Polymerase chain reaction (PCR) technique for simultaneous and practical detection of Listeria monocytogenes (L. monocytogenes) and Escherichia coli O157:H7 (E. coli O157:H7) in the food matrix. EvaGreen-based simplex and duplex real-time PCR showed that specific PCR products were identified by melting curve analysis, and had a reproducible T m of 77.00 6 0.4C for L. monocytogenes and 85.60 6 0.2 for E. coli O157:H7.The absolute detection limit of Eva Green-based simplex and duplex real-time PCR was 0.0001 ng/lL for DNA of both pathogens. The relative detection limit of the simplex and duplex technique was less than 10 cells/mL for the two pathogens. PRACTICAL APPLICATIONSThis study illustrates a rapid, effective and cheap identification method to simultaneously monitor two foodborne pathogens in food samples. The specificity and sensitivity of this method can be used to clearly identify these two foodborne pathogens in practical food samples based on the specific genes of the species. Hence, this detection method is applicable to surveillance measures for these two foodborne pathogens in the food production chain. Moreover, this technique is used by food control laboratories in a secure way.

show abstract

Section: Specificity Of Primersmentioning

confidence: 99%

A Rapid and Cost‐Efficient Technique for Simultaneous/Duplex Detection of Listeria Monocytogenes and Escherichia Coli O157:H7 Using Real Time PCR

Kaynak

Şakalar

2016

Journal of Food Safety

View full text Add to dashboard Cite

show abstract

“…The advantages of the medium are ease of sample preparation, speed and simplicity of process in terms of user input, and the procedure is also far less susceptible to Silicon, silica & its aminosilanization in nanomedicine Review the negative effects of high sample viscosity and particulate density of the mixtures to be separated. It is also less time consuming and cheaper to use, and more convenient for automation [79,80].…”

Section: Silica and Its Surfacementioning

confidence: 99%

Silicon, Silica and its Surface Patterning/Activation with Alkoxy- and Amino-Silanes for Nanomedical Applications

et al. 2011

View full text Add to dashboard Cite

Silica and silicates are widely used in nanomedicine with applications as diverse as medical device coatings to replacement materials in tissue engineering. Although much is known about silica and its synthesis, relatively few biomedical scientists fully appreciate the link that exists between its formulation and its resultant structure and function. This article attempts to provide insight into relevant issues in that context, as well as highlighting their importance in the material's eventual surface patterning/activation with alkoxy- and organo-silanes. The use of aminosilanes in that context is discussed at some length to permit an understanding of the specific variables that are important in the reproducible and robust aminoactivation of surfaces using such molecules. Recent investigative work is cited to underline the fact that although aminosilanization is a historically accepted mechanism for surface activation, there is still much to be explained about how and why the process works in the way it does. In the last section of this article, there is a detailed discussion of two classical approaches for the use of aminosilanized materials in the covalent immobilization of bioligands, amino-aldehyde and amino-carboxyl coupling. In the former case, the use of the homobifunctional coupler glutaraldehyde is explored, and in the latter, carbodiimides. Although these chemistries have long been employed in bioconjugations, it is apparent that there are still variables to be explored in the processes (as witnessed by continuing investigations into the chemistries concerned). Aspects regarding optimization, standardization and reproducibility of the fabrication of amino functionalized surfaces are discussed in detail and illustrated with practical examples to aid the reader in their own studies, in terms of considerations to be taken into account when producing such materials. Finally, the article attempts to remind readers that although the chemistry and materials involved are 'old hat', there is still much to be learnt about the methods involved. The article also reminds readers that although many highly specific and costly conjugation chemistries now exist for bioligands, there still remains a place for these relatively simple and cost-effective approaches in bioligand conjugate fabrication.

show abstract

“…The oligonucleotide array method based on amplification and hybridization of DNA fragments of pathogenic bacteria also takes more than several hours (4). Other methods are performed using expensive special instruments, such as flow cytometry (5) and real time PCR (6). It is imperative to design a rapid, label-free and simple biosensor for the detection of pathogenic bacteria that can be taken to the food source or distribution area.…”

Section: Detection Of Pathogens In the Food Supplymentioning

confidence: 99%

“…The microfluidic chip is placed on top of the prism where it can be imaged by both SPR imaging and epi-fluorescence molecular imaging. (1), two lenses to make a beam expander (2), a polarizer (3), an equilateral prism (4), a long working distance 4X objective, and a lens (6) to focus the image on the CCD (7). The epifluorescence path will image from the top of the sample.…”

Section: Design Of Hybrid Spr/molecular Imaging Prototypementioning

confidence: 99%

A microfluidic-based hybrid SPR/molecular imaging biosensor for the multiplexed detection of foodborne pathogens

et al. 2009

View full text Add to dashboard Cite

It is important to screen our food supply for pathogen contaminations. Current methods to screen for bacterial contamination involve using costly reagents such as antibodies or PCR reagents or time-costly growth in cultures. There is need for portable, real-time, multiplex pathogen detection technology that can predict the safety of food where it is produced or distributed.Surface plasmon resonance (SPR) imaging is a sensitive, label-free method that can detect the binding of an analyte to a surface due to changes in refractive index that occur upon binding. It can be used for label-free detection of the presence of potential pathogens. Simultaneous fluorescence molecular imaging on the other side of the biochip can be used to ascertain pathogen status or functional state which may affect its potential danger to humans or animals.We are designing and testing hybrid microfluidic biochips to detect multiple pathogens using a combination of SPRI and fluorescence imaging. The device consists of an array of gold spots, each functionalized with a peptide targeting a specific pathogen. This peptide biosensor array is enclosed by a PDMS microfluidic flow chamber that delivers a magnetically concentrated sample to be tested. An SPR image is taken from the bottom of the biochip. Image analysis is used to quantify the amount of pathogen (both live and dead) bound to each spot. Since PDMS is very transmissive to visible light, an epi-fluorescence image is taken from the top of the biochip. Fluorescence imaging determines the live:dead ratio of each pathogen using an inexpensive SYTO 9®-Propidium Iodide assay. The volume of sample that the biochip can analyze is small, so possible pathogens are pre-concentrated using immunomagnetic separation. Functionalized magnetic particles are bound to pathogens present in the sample, and a magnet is used to separate them from the bulk fluid.

show abstract

Direct detection of Listeria monocytogenes from milk by magnetic based DNA isolation and PCR

Cited by 71 publications

References 24 publications

A Rapid and Cost‐Efficient Technique for Simultaneous/Duplex Detection of Listeria Monocytogenes and Escherichia Coli O157:H7 Using Real Time PCR

A Rapid and Cost‐Efficient Technique for Simultaneous/Duplex Detection of Listeria Monocytogenes and Escherichia Coli O157:H7 Using Real Time PCR

Silicon, Silica and its Surface Patterning/Activation with Alkoxy- and Amino-Silanes for Nanomedical Applications

A microfluidic-based hybrid SPR/molecular imaging biosensor for the multiplexed detection of foodborne pathogens

Contact Info

Product

Resources

About