Rapid, simple and efficient method for detection of viral genomes on raspberries

Perrin, Aline; Loutreul, Julie; Boudaud, Nicolas; Bertrand, Isabelle; Gantzer, Christophe

doi:10.1016/j.jviromet.2015.08.005

Cited by 24 publications

(22 citation statements)

References 32 publications

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“…The fact that HAV was detected in this study shows that the agent can survive freezing conditions. The fact that there is no control in any production stages to eliminate HAV and it is consumed directly is the reason why these products become the cause of frequent outbreaks (29). These products generally have acidic compositions (pH 2.5-3.3) and contain approximately 5% of sugar.…”

Section: Discussionmentioning

confidence: 99%

Determination of Hepatitis A Virus, Enterobacteriaceae, Coliform and Escherichia coli Contamination of Frozen Raspberries

İnci̇li̇¹

2019

Erciyes Üniversitesi Veteriner Fakültesi Dergisi

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How to cite: İncili GK, Koluman A, Dikici A. Determination of hepatitis A virus, Enterobacteriaceae, coliform and Escherichia coli contamination of frozen raspberries. Erciyes Üniv Vet Fak Derg 2019; 16(1): 44-48.Summary: Hepatitis A is transferred via fecal-oral route, direct contact from person to person or with ingestion of contaminated food. Foods like, raspberries, strawberries and blueberries are important source of hepatitis A virus (HAV). Frozen retail packaged raspberries are analyzed in this study. Frozen raspberry samples have were obtained from local supermarkets monthly between January and December 2015 in Ankara province, Turkey. The samples were evaluated for HAV contamination using ISO/TS 15216-1:2013 in duplicate and Mengovirus was used for process control. Samples that found to be contaminated with HAV were further analyzed for Enterobacteriaceae Coliform and Escherichia coli. Totally 240 samples were analyzed and 20 (8.33%) were found to be contaminated with HAV. The highest HAV prevalence was on 6 th month (25%). On the contrary no HAV was found on 3 rd , 4 th and 10 th months of analyzes. The highest Enterobacteriaceae, Coliform and E. coli counts were found on 2 nd month 4.90, 4.84 and, 3.48 log 10 cfu/g, respectively. The results indicate that even the food relies with the microbiological criteria, there is no guarantee for viral contamination. Additionally, high cost of virus analyzes is another barrier for routine control which has a direct impact on public health. Additional measures like HACCP, GHP and GMP should be applied strictly from farm to fork to avoid contamination with HAV. Dondurulmuş Ahududularda Hepatit A Virüs, Enterobacteriaceae, Koliform ve Escherichia coli Kontaminasyo-nunun Belirlenmesi Özet: Hepatit A virüsü (HAV), genellikle fekal-oral yolla, kişiden kişiye direkt temas veya kontamine gıda ve suyun alınmasıyla bulaşmaktadır. Dondurulmuş ahududu, çilek ve yabanmersini gibi üzümsü meyveler kategorisinde yer alan ürünler, HAV kontaminasyonu açısından önemli kaynaklar arasında yer almaktadır. Bu çalışmada piyasada paketli olarak satılan dondurulmuş ahududu örnekleri kullanılmıştır. Dondurulmuş ahududu örnekleri, 2015 yılında Ankara'da bulunan marketlerden Ocak-Aralık ayları arasında her ay 20 tane olacak şekilde toplanmıştır. Çalışma konusunu oluşturan ahududu örneklerinde HAV analizi ISO/TS 15216-1:2013 uygun olacak şekilde iki paralel olarak yapılmıştır ve proses kontrolü için Mengovirüs kullanılmıştır. HAV pozitif bulunan örneklerde Enterobacteriaceae, Koliform bakteri ve E. coli sayımları gerçekleştirilmiştir. Toplam 240 adet ahududu örneğinin 20 adetinin (%8,33) HAV pozitif olduğu tespit edilmiştir. En yüksek prevalansın 6. ayda (%25) olduğu tespit edilirken, 3, 4 ve 10. aylarda alınan örneklerde ise HAV bulunmamıştır. En yüksek Enterobacteriaceae, koliform ve E. coli sayılarının 2. ayda olduğu ve bu ayda sırasıyla 4,90, 4,84 ve 3,48 log 10 kob/g bakteri olduğu tespit edilmiştir. Bu sonuçlar gıdaların mikrobiyolojik kriterlere göre bakteriyel ajanlar bakımından yeter...

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Section: Discussionmentioning

confidence: 99%

Determination of Hepatitis A Virus, Enterobacteriaceae, Coliform and Escherichia coli Contamination of Frozen Raspberries

İnci̇li̇¹

2019

Erciyes Üniversitesi Veteriner Fakültesi Dergisi

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“…This method can also be applied in other experiments that require detection of a “neutralized” phage, which is merely a phage that is not active against bacteria. Notably, qPCR has already been applied for phage detection in other (than phage neutralizing) conditions: in phage cultures ( Edelman and Barletta, 2003 ; Clokie, 2009 ; Anderson et al, 2011 ; Refardt, 2012 ; Dieterle et al, 2016 ), food ( Imamovic and Muniesa, 2011 ; Flannery et al, 2014 ; Perrin et al, 2015 ; Parente et al, 2016 ; Hartard et al, 2017 ; Muhammed et al, 2017 ), environmental samples ( Farkas et al, 2015 ; Kunze et al, 2015 ; Unnithan et al, 2015 ; Mankiewicz-Boczek et al, 2016 ), and in feces ( Imamovic et al, 2010 ; Chehoud et al, 2016 ), where some interference from antibodies cannot be excluded ( Majewska et al, 2015 ).…”

Section: Discussionmentioning

confidence: 99%

“…Bacteriophages have been quantitatively analyzed and discriminated by real-time qPCR directly in microbiological cultures, and the authors found this method to be a good alternative to the plaque assay ( Edelman and Barletta, 2003 ; Clokie, 2009 ; Anderson et al, 2011 ; Refardt, 2012 ; Dieterle et al, 2016 ). Real-time PCR has been further demonstrated as applicable for a rapid screening allowing phage detection in food (milk, fruits, vegetables, seafood, meat) ( Imamovic and Muniesa, 2011 ; Flannery et al, 2014 ; Perrin et al, 2015 ; Parente et al, 2016 ; Hartard et al, 2017 ) and water samples ( Farkas et al, 2015 ; Kunze et al, 2015 ; Unnithan et al, 2015 ; Mankiewicz-Boczek et al, 2016 ) or in feces ( Imamovic et al, 2010 ; Chehoud et al, 2016 ). However, potential applicability of real-time qPCR for detection of inactivated (non-infective) but still biologically active (e.g., immunoreactive) phage has never been investigated.…”

Section: Introductionmentioning

confidence: 99%

Real-Time qPCR as a Method for Detection of Antibody-Neutralized Phage Particles

et al. 2017

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The most common method for phage quantitation is the plaque assay, which relies on phage ability to infect bacteria. However, non-infective phage particles may preserve other biological properties; specifically, they may enter interactions with the immune system of animals and humans. Here, we demonstrate real-time quantitative polymerase chain reaction (qPCR) detection of bacteriophages as an alternative to the plaque assay. The closely related staphylococcal bacteriophages A3R and 676Z and the coliphage T4 were used as model phages. They were tested in vivo in mice, ex vivo in human sera, and on plastic surfaces designed for ELISAs. T4 phage was injected intravenously into pre-immunized mice. The phage was completely neutralized by specific antibodies within 5 h (0 pfu/ml of serum, as determined by the plaque assay), but it was still detected by qPCR in the amount of approximately 107 pfu/ml of serum. This demonstrates a substantial timelapse between “microbiological disappearance” and true clearance of phage particles from the circulation. In human sera ex vivo, qPCR was also able to detect neutralized phage particles that were not detected by the standard plaque assay. The investigated bacteriophages differed considerably in their ability to immobilize on plastic surfaces: this difference was greater than one order of magnitude, as shown by qPCR of phage recovered from plastic plates. The ELISA did not detect differences in phage binding to plates. Major limitations of qPCR are possible inhibitors of the PCR reaction or free phage DNA, which need to be considered in procedures of phage sample preparation for qPCR testing. We propose that phage pharmacokinetic and pharmacodynamic studies should not rely merely on detection of antibacterial activity of a phage. Real-time qPCR can be an alternative for phage detection, especially in immunological studies of bacteriophages. It can also be useful for studies of phage-based drug nanocarriers or biosensors.

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“…virus elution and concentration from various matrices which allow a high recovery, needs to be addressed. Direct extraction of RNA from berry surfaces by immersion into lysis buffer was efficient in detecting some NoV surrogates on artificially contaminated berries (Perrin et al, 2015). A further step towards complete validation, however, requires demonstrated detection of viral pathogens in naturally contaminated samples and comparison of performance between laboratories.…”

Section: Iso/cen Methodsmentioning

confidence: 99%

Foodborne viruses: Detection, risk assessment, and control options in food processing

Bosch

Gkogka

Guyader

et al. 2018

International Journal of Food Microbiology

206

140

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In a recent report by risk assessment experts on the identification of food safety priorities using the Delphi technique, foodborne viruses were recognized among the top rated food safety priorities and have become a greater concern to the food industry over the past few years. Food safety experts agreed that control measures for viruses throughout the food chain are required. However, much still needs to be understood with regard to the effectiveness of these controls and how to properly validate their performance, whether it is personal hygiene of food handlers or the effects of processing of at risk foods or the interpretation and action required on positive virus test result. This manuscript provides a description of foodborne viruses and their characteristics, their responses to stress and technologies developed for viral detection and control. In addition, the gaps in knowledge and understanding, and future perspectives on the application of viral detection and control strategies for the food industry, along with suggestions on how the food industry could implement effective control strategies for viruses in foods. The current state of the science on epidemiology, public health burden, risk assessment and management options for viruses in food processing environments will be highlighted in this review.

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Rapid, simple and efficient method for detection of viral genomes on raspberries

Cited by 24 publications

References 32 publications

Determination of Hepatitis A Virus, Enterobacteriaceae, Coliform and Escherichia coli Contamination of Frozen Raspberries

Determination of Hepatitis A Virus, Enterobacteriaceae, Coliform and Escherichia coli Contamination of Frozen Raspberries

Real-Time qPCR as a Method for Detection of Antibody-Neutralized Phage Particles

Foodborne viruses: Detection, risk assessment, and control options in food processing

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