Differential detection of pathogenic Yersinia spp. by fluorescence in situ hybridization

Rohde, Alexander; Hammerl, Jens A.; Appel, Bernd; Dieckmann, Ralf; Dahouk, Sascha Al

doi:10.1016/j.fm.2016.09.013

Cited by 18 publications

(10 citation statements)

References 27 publications

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“…However, discrimination of the two organisms was possible using a fluorescently labeled competitor containing a locked nucleic acid (LNA) targeting Y. pseudotuberculosis that specifically detected Y. pseudotuberculosis, but not Y. pestis (S2 Fig). The incorporation of LNA-bases in FISH-probes has also been reported to increase the stringency of FISH-probes in other studies [27,50,51]. Urea-based hybridizations exhibited specificity identical to formamide-based FISH.…”

Section: Plos Onementioning

confidence: 79%

“…However, FISH was shown to perform better than most biosensors for the detection of B. anthracis spores from air [23]. Another recent study showed that in minced meat, after a short enrichment step, one colony forming unit/gram of Y. enterocolitica was detectable by FISH, making it more sensitive than currently used ISO methods in food microbiology [27].…”

Section: Plos Onementioning

confidence: 99%

“…However, these studies all relied on parallel or subsequent hybridizations with a maximum of three probes per reaction in order to identify the relevant bacteria. The causative agents of anthrax, plague, tularemia and brucellosis have already been targeted by FISH in previous studies [23][24][25][26][27][28], but a comprehensive probe set for multiplexed in situ detection of a wide range of biodefense relevant bacteria has hitherto not been established. The situation is further complicated by the fact that first preliminary tests for biological agents might have to be performed on site in deployed field laboratories.…”

Section: Introductionmentioning

confidence: 99%

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NOTIFy (non-toxic lyophilized field)-FISH for the identification of biological agents by Fluorescence in situ Hybridization

et al. 2020

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The rapid and reliable diagnostics of highly pathogenic bacteria under restricted field conditions poses one of the major challenges to medical biodefense, especially since false positive or false negative reports might have far-reaching consequences. Fluorescence in situ hybridization (FISH) has the potential to represent a powerful microscopy-based addition to the existing molecular-based diagnostic toolbox. In this study, we developed a set of FISHprobes for the fast, matrix independent and simultaneous detection of thirteen highly pathogenic bacteria in different environmental and clinical sample matrices. Furthermore, we substituted formamide, a routinely used chemical that is toxic and volatile, by non-toxic urea. This will facilitate the application of FISH under resource limited field laboratory conditions. We demonstrate that hybridizations performed with urea show the same specificity and comparable signal intensities for the FISH-probes used in this study. To further simplify the use of FISH in the field, we lyophilized the reagents needed for FISH. The signal intensities obtained with these lyophilized reagents are comparable to freshly prepared reagents even after storage for a month at room temperature. Finally, we show that by the use of nontoxic lyophilized field (NOTIFy)-FISH, specific detection of microorganisms with simple and easily transportable equipment is possible in the field.

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Section: Plos Onementioning

confidence: 79%

Section: Plos Onementioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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NOTIFy (non-toxic lyophilized field)-FISH for the identification of biological agents by Fluorescence in situ Hybridization

et al. 2020

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“…The FISH method is a useful tool for the initial identification of microbial pathogens, and it is considered quicker and associated with less hands-on time than traditional culture methods [ 32 , 50 ]. However, this technique on its own cannot ensure that bacteria remain alive.…”

Section: Discussionmentioning

confidence: 99%

Combination of Direct Viable Count and Fluorescent In Situ Hybridization (DVC-FISH) as a Potential Method for Identifying Viable Vibrio parahaemolyticus in Oysters and Mussels

García‐Hernández

Cedillo-Hernandez

Moreno

2021

Foods

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Vibrio parahaemolyticus is a human food-borne pathogen with the ability to enter the food chain. It is able to acquire a viable, non-cultivable state (VBNC), which is not detected by traditional methods. The combination of the direct viable count method and a fluorescent in situ hybridization technique (DVC-FISH) makes it possible to detect microorganisms that can present VBNC forms in complex samples The optimization of the in vitro DVC-FISH technique for V. parahaemolyticus was carried out. The selected antibiotic was ciprofloxacin at a concentration of 0.75 μg/mL with an incubation time in DVC broth of 5 h. The DVC-FISH technique and the traditional plate culture were applied to detect and quantify the viable cells of the affected pathogen in artificially contaminated food matrices at different temperatures. The results obtained showed that low temperatures produced an important logarithmic decrease of V. parahaemolyticus, while at 22 °C, it proliferated rapidly. The DVC-FISH technique proved to be a useful tool for the detection and quantification of V. parahaemolyticus in the two seafood matrices of oysters and mussels. This is the first study in which this technique has been developed to detect viable cells for this microorganism.

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“…in pork using LNA probe in 2017. 24 PNA and LNA are unnatural molecules, and they are not substrates for enzymes that modify DNA, RNA and proteins. Therefore, much more effective than DNA or RNA in FISH detection.…”

Section: Locked Nucleic Acids (Lna)mentioning

confidence: 99%

Fluorescence in situ hybridization (FISH) in food pathogen detection

Pgi

Rmusk

2018

IJMBOA

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Diseases due to food borne microbial pathogens are major public health concern in the world. Different methods are used to detect food borne pathogens from different types of foods to prevent from these diseases. Out of those methods, molecular detection methods have received more concern in the recent past. Fluorescence in situ Hybridization is one of such methods used in the detection of food borne microbial pathogens. The present situation of using Fluorescence in situ Hybridization to detect food borne microbial pathogens is review in this article.

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Differential detection of pathogenic Yersinia spp. by fluorescence in situ hybridization

Cited by 18 publications

References 27 publications

NOTIFy (non-toxic lyophilized field)-FISH for the identification of biological agents by Fluorescence in situ Hybridization

NOTIFy (non-toxic lyophilized field)-FISH for the identification of biological agents by Fluorescence in situ Hybridization

Combination of Direct Viable Count and Fluorescent In Situ Hybridization (DVC-FISH) as a Potential Method for Identifying Viable Vibrio parahaemolyticus in Oysters and Mussels

Fluorescence in situ hybridization (FISH) in food pathogen detection

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