Multiplex real-time PCRs for detection of Salmonella, Listeria monocytogenes, and verotoxigenic Escherichia coli in carcasses of slaughtered animals

Denis, E.; Bielińska, Katarzyna; Wieczorek, Kinga; Osek, Jacek

doi:10.1515/jvetres-2016-0044

Cited by 12 publications

(6 citation statements)

References 25 publications

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“…7,10,32,44 One fetal brain sample (case 5) was also tested for Listeria by PCR assay. 13 PCR for Chlamydia spp. was performed on fresh tissue from 18 animals (2 cows, 1 calf, 15 fetuses) that included brain only (1 cow, 1 calf, 8 fetuses), brain and pericardium (1 fetus), lung (1 cow), and pooled samples of brain, spleen, lung (1 fetus) or brain, lung, liver, and kidney (2 fetuses) or lung, liver, and kidney (3 fetuses).…”

Section: Molecular Diagnostic Testing For Abortogenic Agentsmentioning

confidence: 99%

Meningoencephalitis, Vasculitis, and Abortions Caused by Chlamydia pecorum in a Herd of Cattle

Struthers

Lim²,

Ferguson

et al. 2021

Vet Pathol

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A cow dairy ( n = 2000) in close proximity to a sheep flock had third-trimester abortions and fatalities in cows and calves over a 14-month period. Eighteen of 33 aborted fetuses (55%) had multifocal random suppurative or mononuclear meningoencephalitis with vasculitis. Seventeen of these affected fetuses had intracytoplasmic bacteria in endothelial cells, and 1 fetus with pericarditis had similar bacteria within mesothelial cells or macrophages. Immunohistochemistry for Chlamydia spp. or polymerase chain reaction (PCR) for Chlamydia pecorum or both, performed on brain or pooled tissue, were positive in all 14 tested fetuses that had meningoencephalitis and in 4/4 calves and in 3/4 tested cows that had meningoencephalitis and thrombotic vasculitis. In 1 calf and 11/11 fetuses, C. pecorum PCR amplicon sequences were 100% homologous to published C. pecorum sequences. Enzootic chlamydiosis due to C. pecorum was the identified cause of the late term abortions and the vasculitis and meningoencephalitis in fetuses, calves, and cows. C. pecorum, an uncommon bovine abortogenic agent, is a differential diagnosis in late-term aborted fetuses with meningoencephalitis, vasculitis, and polyserositis.

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Section: Molecular Diagnostic Testing For Abortogenic Agentsmentioning

confidence: 99%

Meningoencephalitis, Vasculitis, and Abortions Caused by Chlamydia pecorum in a Herd of Cattle

Struthers

Lim²,

Ferguson

et al. 2021

Vet Pathol

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“…Traditional bacteriological methods are, however, time-consuming because they may last up to a few days ( 1 , 8 , 22 ). L. monocytogenes , being a pathogen of humans and animals, requires faster and less laborious diagnostic methods such as real-time PCR ( 2 , 5 ). For detection of real-time PCR products non-specific dyes and specific fluorophore-labelled oligonucleotide probes are used.…”

Section: Discussionmentioning

confidence: 99%

Detection of Listeria spp. and Listeria monocytogenes in biological samples by SYBR Green I and TaqMan probe-based real-time PCRs

Kędrak-Jabłońska

Budniak

Krupa

et al. 2017

Journal of Veterinary Research

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IntroductionThe aim of the study was the application and comparison of real-time PCR methods based on the fluorescence of SYBR Green I intercalating dye and TaqMan probes for the detection of the 23S rDNA gene of Listeria spp. and the hlyA gene of Listeria monocytogenes in biological samples of the liver, brain, and blood.Material and MethodsFive strains of L. monocytogenes and single strains of each species L. ivanovii, L. innocua,L. grayi, L. welshimeri, and L. seeligeri were used for the experiments. Additionally, five strains of other species of bacteria were used for evaluation of the specificity of tests. In the first stage of the study SYBR Green I real-time PCRs, one allowing detection of the 23S rDNA gene and two based on the amplification the hlyA gene, were performed. In the next part, three TaqMan probe-based real-time PCRs allowing confirmation of belonging to Listeria spp. and L. monocytogenes were conducted.ResultsThe observation of amplification curves in real-time PCRs enabled the detection of both genes. A high regression coefficient of 0.99 was found for all reactions. Specific amplification products were obtained for the 23S rDNA and hlyA genes which confirm their belonging to Listeria spp. and L. monocytogenes, respectively. Other microbial species did not reveal real-time PCR products.ConclusionBoth real-time PCR methods for the detection of Listeria spp. and L. monocytogenes in biological samples demonstrated a significant sensitivity and high specificity.

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“…Compared to conventional PCR, real-time PCR offers better options for multiplexing several targets in a single reaction. Recently, there has been an increase in the development of real-time multiplex PCRs for the detection of multiple pathogens including foodborne pathogens (41)(42)(43)(44)(45)(46). In one such study, Hu et al developed a molecular beaconbased multiplex real-time PCR for the detection of various foodborne pathogens, namely S. enterica subsp.…”

Section: Real-time Pcrmentioning

confidence: 99%

Molecular Tools To Study Preharvest Food Safety Challenges

Kumar

Thakur

2018

Microbiol Spectr

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Preharvest food safety research and activities have advanced over time with the recognition of the importance and complicated nature of the preharvest phase of food production. In developed nations, implementation of preharvest food safety procedures along with strict monitoring and containment at various postharvest stages such as slaughter, processing, storage, and distribution have remarkably reduced the burden of foodborne pathogens in humans. Early detection and adequate surveillance of pathogens at the preharvest stage is of the utmost importance to ensure a safe meat supply. There is an urgent need to develop rapid, cost-effective, and point-of-care diagnostics which could be used at the preharvest stage and would complement postmortem and other quality checks performed at the postharvest stage. With newer methods and technologies, more efforts need to be directed toward developing rapid, sensitive, and specific methods for detection or screening of foodborne pathogens at the preharvest stage. In this review, we will discuss the molecular methods available for detection and molecular typing of bacterial foodborne pathogens at the farm. Such methods include conventional techniques such as endpoint PCR, real-time PCR, DNA microarray, and more advanced techniques such as matrix-assisted layer desorption ionization-time of flight mass spectrometry and whole-genome sequencing.

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Multiplex real-time PCRs for detection of Salmonella, Listeria monocytogenes, and verotoxigenic Escherichia coli in carcasses of slaughtered animals

Cited by 12 publications

References 25 publications

Meningoencephalitis, Vasculitis, and Abortions Caused by Chlamydia pecorum in a Herd of Cattle

Meningoencephalitis, Vasculitis, and Abortions Caused by Chlamydia pecorum in a Herd of Cattle

Detection of Listeria spp. and Listeria monocytogenes in biological samples by SYBR Green I and TaqMan probe-based real-time PCRs

Molecular Tools To Study Preharvest Food Safety Challenges

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