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Organization and ELISA-Based Results of the First Proficiency Testing to Evaluate the Ability of European Union Laboratories to Detect Staphylococcal Enterotoxin Type B (SEB) in Buffer and Milk

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ABSTRACT

The aim of this work was to organize the first proficiency test (PT) dedicated to staphylococcal enterotoxin B (SEB) detection in milk and buffer solutions. This paper describes the organization of the PT trial according to EN ISO 17043 requirements. Characterization of the SEB stock solution was performed using SDS-PAGE and SE-specific ELISA, and amino acid analysis was used to assign its protein concentration. The solution was then used to prepare six PT materials (four milk and two buffer batches) at a ng/g toxin level, which included one blank and one SEA-containing milk as specificity control. Suitable material homogeneity and stability were assessed using screening and quantitative ELISAs. Among the methods used by the participants, ELISA-based methods demonstrated their efficiency for the detection of SEB in both simple and complex matrices. The results serve as a basis for further improving the detection capabilities in expert laboratories and can therefore be considered as a contribution to biopreparedness.

No MeSH data available.


Stability data obtained for the milk and buffer samples spiked by SEB. Data were presented as recovery calculated from the measured SEB concentrations in stability samples and the assigned value corresponding to the homogeneity value.
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toxins-08-00268-f003: Stability data obtained for the milk and buffer samples spiked by SEB. Data were presented as recovery calculated from the measured SEB concentrations in stability samples and the assigned value corresponding to the homogeneity value.

Mentions: The homogeneity test was performed on 20 samples one week before samples dispatch. In order to assess to stability of SEB in each batch (M1 to M4, B1 and B2), six randomly selected samples were analyzed after receiving participant results (six weeks after dispatching). Then, these six replicates were compared to the mean of the 20 values obtained in the homogeneity study (Equation (1)). Results are presented in Figure 3.(1)[Buffer 1 recovery]%=SEB concentration measured in Buffer 1 (stability)the assigned value for Buffer 1 (homogeneity)×100


Organization and ELISA-Based Results of the First Proficiency Testing to Evaluate the Ability of European Union Laboratories to Detect Staphylococcal Enterotoxin Type B (SEB) in Buffer and Milk
Stability data obtained for the milk and buffer samples spiked by SEB. Data were presented as recovery calculated from the measured SEB concentrations in stability samples and the assigned value corresponding to the homogeneity value.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5037494&req=5

toxins-08-00268-f003: Stability data obtained for the milk and buffer samples spiked by SEB. Data were presented as recovery calculated from the measured SEB concentrations in stability samples and the assigned value corresponding to the homogeneity value.
Mentions: The homogeneity test was performed on 20 samples one week before samples dispatch. In order to assess to stability of SEB in each batch (M1 to M4, B1 and B2), six randomly selected samples were analyzed after receiving participant results (six weeks after dispatching). Then, these six replicates were compared to the mean of the 20 values obtained in the homogeneity study (Equation (1)). Results are presented in Figure 3.(1)[Buffer 1 recovery]%=SEB concentration measured in Buffer 1 (stability)the assigned value for Buffer 1 (homogeneity)×100

View Article: PubMed Central - PubMed

ABSTRACT

The aim of this work was to organize the first proficiency test (PT) dedicated to staphylococcal enterotoxin B (SEB) detection in milk and buffer solutions. This paper describes the organization of the PT trial according to EN ISO 17043 requirements. Characterization of the SEB stock solution was performed using SDS-PAGE and SE-specific ELISA, and amino acid analysis was used to assign its protein concentration. The solution was then used to prepare six PT materials (four milk and two buffer batches) at a ng/g toxin level, which included one blank and one SEA-containing milk as specificity control. Suitable material homogeneity and stability were assessed using screening and quantitative ELISAs. Among the methods used by the participants, ELISA-based methods demonstrated their efficiency for the detection of SEB in both simple and complex matrices. The results serve as a basis for further improving the detection capabilities in expert laboratories and can therefore be considered as a contribution to biopreparedness.

No MeSH data available.