A técnica da fluorescência amarelo-esverdeada brilhante (BGYF) como método de triagem na avaliação de milho eventualmente contaminado com aflatoxina, utilizada por uma indústria alimentícia do Estado de São Paulo em 61 amostras de milho, foi comparada com a técnica BGYF padronizada, aplicada em laboratório antes e após trituração dos grãos e, também, com a técnica da cromatografia em camada delgada (CCD). Os resultados da indústria geraram menor número de resultados falso-positivos do que a técnica BGYF padronizada. Por outro lado, apenas os resultados da indústria apresentaram amostras falso-negativas, ao contrário da BGYF padronizada, que não as apresentou. Conclui-se que existe uma inadequação do número de pontos fluorescentes adotados como critério de rejeição em ambas as técnicas, devendo-se proceder estudos, com maior número de amostras, para determinar o número adequado de pontos fluorescentes a ser adotado como critério além de outros fatores que possam influenciar a metodologia.
The use of antibodies in immunodiagnostic kits generally implies the conjugation of these proteins with other molecules such as chromophores or fluorochromes. The development of more sensitive quality control procedures than spectrophotometry is essential to assure the use of better fluorescent conjugates since the fluorescent conjugates are critical reagents for a variety of immunodiagnostic kits. In this article, we demonstrate a new flow cytometric protocol to evaluate conjugates by molecules of equivalent soluble fluorochromes (MESF) and by traditional flow cytometric analysis. We have coupled microspheres with anti-IgG-PE and anti-HBSAg-PE conjugates from distinct manufactures and/or different lots and evaluated by flow cytometry. Their fluorescence intensities were followed for a period of 18 months. Our results showed that there was a great difference in the fluorescence intensities between the conjugates studied. The differences were observed between manufactures and lots from both anti-IgG-PE and anti-HBSAg-PE conjugates. Coefficients of variation (CVs) showed that this parameter can be used to determine better coupling conditions, such as homogenous coupling. The MESF analysis, as well as geometric mean evaluation by traditional flow cytometry, showed a decrease in the values for all conjugates during the study and were indispensable tools to validate the results of stability tests. Our data demonstrated the feasibility of the flow cytometric method as a standard quality control of immunoassay kits.
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