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Tsuchiura, Japan

Kawasaki S.,Japan National Food Research Institute | Kusano K.,Kirin Holdings Company | Arai R.,Kirin Holdings Company | Komeda T.,Kirin Holdings Company | And 2 more authors.
Journal of Food, Agriculture and Environment | Year: 2012

A multiplex (m-)PCR method was developed for simultaneous detection of Salmonella spp., Listeria monocytogenes, Escherichia coli O157:H7 and Staphylococcus aureus in processed foods. The m-PCR detection protocol consisted of three steps: 1) 20-h pre-enrichment culturing, 2) simple DNA extraction, and 3) m-PCR detection. For pre-enrichment of pathogens from food samples, [TA10] simultaneous growth broth was used. [TA10] broth does not contain selective agents and is of practical use for these four pathogenic bacteria without modification. For m-PCR detection, the targeted genes included a Salmonella species-specific sequence within a 1.8-kb HindIII DNA fragment, L. monocytogenes hlyA, E. coli O157:H7 eaeA, and S. aureus femA. The m-PCR assay with enrichment broth was evaluated in 13 types of spiked food samples (including pasteurized milk, yogurt, processed cheese, etc.), and targeted genes specific for each pathogen simultaneously. Assay sensitivity was 10° CFU for each pathogen in 25 g of sample with a 20-h enrichment period. This m-PCR method is reliable and useful for rapid screening of processed food samples contaminated with Salmonella spp., L. monocytogenes, E. coli O157:H7 or S. aureus.


Patent
Prima Meat Packers Ltd. | Date: 2010-02-23

A method is disclosed which rapidly detect allergens with good accuracy by efficiently extracting the allergens from a test sample such as food containing various allergens and eliminating non-specific reactions accompanying the disintegration of colloidal gold conjugated to antibodies by not using a reducing agent. The method uses a developing solution containing at least 10% by weight of FBS in an immunochromatography method which comprises using a colloidal gold-labeled antibody in which a colloidal gold is bound to a monoclonal antibody against denatured and native allergen, a development support wherein a monoclonal antibody against denatured and native allergen recognizing an epitope different from that recognized by the colloidal gold-labeled antibody is fixed, measurement samples of the allergens extracted from the test sample with an anionic surfactant such as SDS and a thiosulfate or an anionic surfactant such as SDS and a non-ionic surfactant such as Tween 20, and the developing solution; developing the developing solution on the development support; and then detecting the allergens based on the presence of the deposition of the colloidal gold.


Kato S.,Prima Meat Packers Ltd. | Yagi T.,Prima Meat Packers Ltd. | Kato A.,Prima Meat Packers Ltd. | Yamamoto S.,Prima Meat Packers Ltd. | And 2 more authors.
Journal of AOAC International | Year: 2015

The labeling of seven specific allergenic ingredients (egg, milk, wheat, buckwheat, peanut, shrimp, and crab) is mandatory in Japan. To ensure proper labeling, two kinds of ELISA kits using polyclonal antibodies have been developed. However, we developed two novel ELISA kits using monoclonal antibodies with improved specificity, the Allergeneye ELISA Egg (AE-Egg) and Allergeneye ELISA Milk (AE-Milk) Kits, to detect egg and milk proteins in processed foods, respectively. Five types of processed food containing 10 mg/kg of egg or milk soluble protein were prepared for an interlaboratory study of the performance of these kits. The kits showed a relatively high reproducibility level of interlaboratory precision (AE-Egg RSDR, 3.7-5.7%; AE-Milk RSDR, 6.8-10.5%) and satisfied the recovery rate stipulated by Japanese guidelines (AE-Egg, 61.6-89.3%; AE-Milk, 52.1-67%) for all processed foods. Our results suggest that the AE-Egg and AE-Milk Kits are precise and reliable tools for detecting egg or milk proteins in processed foods.


Kamisaki-Horikoshi N.,Prima Meat Packers Ltd. | Okada Y.,Prima Meat Packers Ltd. | Takeshita K.,Prima Meat Packers Ltd. | Sameshima T.,Prima Meat Packers Ltd. | Arihara K.,Kitasato University
Nippon Shokuhin Kagaku Kogaku Kaishi | Year: 2012

In order to clarify the factors affecting the growth rate of Listeria monocytogenes in meat products, L. monocytogenes was inoculated into various meat products and raw ham, which is served unheated, and its growth rate along with the effects of pH and water activity (aW) on the growth rate were investigated. L. monocytogenes grew in chicken nuggets and cooked pork loin ham during storage for 60 days at 10°C but did not grow in Vienna sausage which contains sorbic acid over the same period. The effect of sorbic acid on growth rate of L. monocytogenes should be investigated in more detail. On the other hand, L. monocytogenes did not grow in raw ham with pH≤6.0 and a W≤0.93 during storage for 60 days at 10°C. These results suggest that the risk of listeriosis in raw ham can be reduced by adjusting pHand aW.


Kato S.,Prima Meat Packers Ltd. | Yagi T.,Prima Meat Packers Ltd. | Namioka M.,Prima Meat Packers Ltd. | Akimoto M.,Prima Meat Packers Ltd. | Arihara K.,Kitasato University
Nippon Shokuhin Kagaku Kogaku Kaishi | Year: 2014

We developed a novel ELISA kit for cow's milk detection in accordance with the official Japanese method for extraction (official extraction method), which is used for monitoring the appropriateness of food product labeling in Japan. The monoclonal antibody used in the kit was prepared using native or reduced and carboxymethylated cow's milk β -lactoglobulin as an immunizing antigen. The newly developed ELISA kit detected cow' s milk protein solubilized in 2-mercaptoethanol and sodium dodecyl sulfate, which is used in the official extraction method. The detectable range of egg protein concentrations was 0. 8-50. 0 ng/mL (equivalent to 0. 3-20. 0 μg/g food). Crossreactivity with both sheep's milk and duck's milk, but not with other food ingredients, was observed. Spike recovery tests using model processed foods showed that recovery ranged from 61. 1-77. 9%, and intra- and inter-assay coefficients of variation were less than 6.5% and 7.9%, respectively. Based on the official extraction method for quantitative analysis of cow's milk in specified food ingredients, these results suggest that a simple, monoclonal antibody-based ELISA kit is as effective as the conventional method for cow's milk protein detection. © 2014, Japanese Society for Food Science and Technology.

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