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Halbmayr-Jech E.,Romer Labs Division Holding GmbH | Rogers A.,Romer Labs UK Ltd. | Don C.,Foodphysica | Prinster M.,Romer Labs Inc.
Journal of AOAC International | Year: 2015

The Protein and Enzymes Technical Committee of American Association of Cereal Chemists initiated a collaborative study to confirm whether the G12 antibody-based sandwich ELISA test kit is able to detect gluten in the lower mg/kg (ppm) level. Twenty laboratories investigated 24 heat-treated and non-heat-treated blind-coded samples with incurred gluten levels up to 100 mg/kg. The method has been validated for testing foods to conform to the defined Codex thresholds for gluten in gluten-free products at less than 20 mg gluten/kg. The collaborative study showed that low levels of gluten could be detected by G12 Sandwich ELISA with reproducibility RSDR of 32% and repeatability RSDr of 16%. Incurred samples showed a recovery between 62 and 135%. It is recommended that the method be accepted by AOAC as Official First Action.

Wight W.D.,Michigan State University | Labuda R.,Romer Labs Division Holding GmbH | Walton J.D.,Michigan State University
BMC Microbiology | Year: 2013

Background: HC-toxin, a cyclic tetrapeptide, is a virulence determinant for the plant pathogenic fungus Cochliobolus carbonum. It was recently discovered that another fungus, Alternaria jesenskae, also produces HC-toxin. Results: The major genes (collectively known as AjTOX2) involved in the biosynthesis of HC-toxin were identified from A. jesenskae by genomic sequencing. The encoded orthologous proteins share 75-85% amino acid identity, and the genes for HC-toxin biosynthesis are duplicated in both fungi. The genomic organization of the genes in the two fungi show a similar but not identical partial clustering arrangement. A set of representative housekeeping proteins show a similar high level of amino acid identity between C. carbonum and A. jesenskae, which is consistent with the close relatedness of these two genera within the family Pleosporaceae (Dothideomycetes). Conclusions: This is the first report that the plant virulence factor HC-toxin is made by an organism other than C. carbonum. The genes may have moved by horizontal transfer between the two species, but it cannot be excluded that they were present in a common ancestor and lost from other species of Alternaria and Cochliobolus. © 2013 Wight et al.; licensee BioMed Central Ltd.

Javorekova S.,Slovak University of Agriculture | Labuda R.,Romer Labs Division Holding GmbH | Makova J.,Slovak University of Agriculture | Novak J.,Slovak University of Agriculture | And 2 more authors.
Mycopathologia | Year: 2012

A total of 939 isolates of 11 genera representing 15 species of keratinophilic fungi were isolated and identified from the soils of three long-term fold-grazed pastures in national parks of Slovakia (Pod Ploskou, Strungový príslop, and Pod Kečkou) and one non-fold-grazed pasture in sierra Stolicke vrchy (Diel) using the hair-baiting technique. Keratinophilic fungi were present in all soil samples with a prevalence of Trichophyton ajelloi and Paecilomyces lilacinus. These fungi were more abundant in soil from fold-grazed pasture (Strungový príslop) compared to non-fold-grazed pasture (Diel). The occurrence of the other keratinophilic fungi was substantially lower, likely because of low pH in some soils. © 2012 Springer Science+Business Media B.V.

Goh P.H.,Romer Labs Singapore Pte Ltd | Zheng M.,Romer Labs Singapore Pte Ltd | Cvak B.,Romer Labs Division Holding GmbH
Asian Pacific Journal of Tropical Disease | Year: 2014

Introduction: Aflatoxins (B1, B2, G1 and G2) are toxic secondary metabolites of, primarily, the fungi Aspergillus flavus and Aspergillus parasiticus. Aflatoxins are potent carcinogens which cause liver diseases in livestock, domestic animals and humans throughout the world. Regulations for aflatoxins in commodities and food exist in more than 100 countries. A rapid quantitative lateral flow test, namely "Agrastrip", was developed to measure total aflatoxin levels in maize. The test is developed based on the competitive immuno-chromatography principle. The total assay time for a sample extract is approximately 5 minutes. The results from the test are comparable to the reference HPLC or LC-MS/MS methods. Objective: To validate the quantitative AgraStrip total aflatoxin test in maize samples. Methods: Aflatoxins were extracted from ground maize sample using 70% methanol. To start an assay, buffer was added into a microwell to dissolve the pre-coated antibody-colloidal gold conjugate particle in the microwell. This was followed by mixing the dissolved conjugate with sample extract. A test strip was then placed into the mixture in microwell and incubated in a heat block incubator for 3 minutes. The test line zone in the test strip is able to capture the conjugate and form a visible line. The concentration of aflatoxins present in the sample is inversely proportional to the colour intensity of the test line. A line is always visible in positive control zone regardless of the presence of aflatoxins. The test strip is analyzed by the AgraVision® Reader and concentration of aflatoxins is determined. Results & Discussion: The test was able to measure aflatoxins in the range of 5-100μg/kg. Limit of detection for the test kit on corn sample was 3.6μg/kg. Accuracy and precision of the test on maize sample naturally contaminated with aflatoxins were comparable to the results form reference HPLC or LC-MS/MS methods. The shelf-life of the test kit was 12 months. Conclusion: Agrastrip total aflatoxin test is able to screen total aflatoxins in maize samples in its quantitation range. © 2014 Asian Pacific Tropical Medicine Press.

Mogensen J.M.,Technical University of Denmark | Moller K.A.,Technical University of Denmark | Von Freiesleben P.,Technical University of Denmark | Labuda R.,Romer Labs Division Holding GmbH | And 6 more authors.
Journal of Industrial Microbiology and Biotechnology | Year: 2011

Tolypocladium inflatum is known primarily for its production of the cyclosporines that are used as an immunosuppressive drug. However, we report here the production of the carcinogenic fumonisins B2 and B 4 by this biotechnologically relevant fungal genus. These mycotoxins were detected in 11 strains tested from three species: Tolypocladium inflatum, T. cylindrosporum, and T. geodes. Production of fumonisins by Fusarium spp. and Aspergillus niger is highly medium- and temperature-dependent, so the effect of these parameters on fumonisin production by three T. inflatum strains was studied. Maximum production was achieved on media with high sugar content incubated at 25-30°C. Since these results demonstrate that fumonisin production could be widespread within the genus Tolypocladium, the potential contamination of commercial cyclosporine preparations with fumonisins needs to be investigated. © 2010 Society for Industrial Microbiology.

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