Abstract
The ability of viruses to persist in the environment and on fresh produce, as well as their low infectious doses, allows even a small amount of contamination to cause foodborne viral outbreaks. Human Adenoviruses (hAdVs) of subgroup F (enteric serotypes 40 and 41) are known to be associated with 5–20% worldwide of acute gastroenteritis cases among infants and young children. The present study aimed to evaluate a simple and cost-effective, one-step, single-tube adenovirus type 40/41 specific loop-mediated isothermal amplification (LAMP) assay for the detection of hAdV40/41 DNA in a series of ready-to-eat food samples (strawberries, sour cherries, lettuce, cherry tomatoes and green onions). No LAMP assay has previously been reported for the detection of these virus types in food samples. Results were obtained within 60 min, under isothermal conditions at 69°C. After amplification, the products were detected either by observing a ladder pattern following gel electrophoresis, or a color change with the addition of SYBR Green I to the reaction tube. The limits of detection of hAdV40/41 in food samples were found to be >30 GCs per reaction, when a nucleic acids extraction step was included in the analysis. A 1 log higher (>100 GCs/reaction) detection limit was found for lettuce, cherry tomatoes and green onions, without a nucleic acids extraction step included before the isothermal amplification. The LAMP assay for the virological analysis of food samples is expected to provide a robust, innovative, powerful, cheap and fast monitoring tool, without the need of sophisticated equipment, which will be available for food safety testing by the food industry and the public food health authorities.
Highlights
Ready-to-eat (RTE) fruits and vegetables are considered important components of a healthy and balanced diet and recognized as an important source of nutrients for humans (Meng and Gerba, 1996, Nuanualsuwan and Oliver, 2003a)
The aim of the present study was the evaluation of a simple, cost-effective, onestep, single-tube adenovirus types 40/41 specific loop-mediated isothermal amplification (LAMP) assay, which has been recently published by our group (Ziros et al, 2015), for the detection of hAdV40/41 in a series of fresh ready-toeat (RTE) food samples
LAMP specificity The specific isothermal amplification of the DNA of hAdV40/41 strains on food samples generated ladder-like pattern bands on agarose gel
Summary
Ready-to-eat (RTE) fruits and vegetables are considered important components of a healthy and balanced diet and recognized as an important source of nutrients for humans (Meng and Gerba, 1996, Nuanualsuwan and Oliver, 2003a). The majority of adenoviruses cause respiratory tract diseases, some of them are associated with gastroenteritis (Van et al, 1992). HAdVs species F types 40 and 41 are enteric pathogens which are associated with 5–20% of acute gastroenteritis cases among infants and young children (Haramoto et al, 2007, Bányai et al, 2009, Haramoto et al, 2010, Sdiri-Loulizi et al, 2009, Lee et al, 2012, RodríguezLázaro et al, 2012). In 2000, Notomi et al developed a novel nucleic acid amplification method, designated loop-mediated isothermal amplification (LAMP) This novel assay is advantageous since it has high specificity, sensitivity, low cost and it can be used as an easy diagnostic tool without the need of sophisticated equipment, by the food industry and the Public health authorities (Notomi et al, 2000)
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