Validación del método analítico para diagnosticar Botrytis cinerea mediante el sistema de identificación microbiana BIOLOG, y el aseguramiento de la calidad del ensayo mediante análisis genético

Abstract

AGROCALIDAD is the Agency in charge of the regulation and control of the agricultural sanitation and food safety in primary production. To guarantee the quality and safety of agricultural production, it implements plans, programs and projects for plant sanitation that support trade flows and contribute to food sovereignty (AGROCALIDAD, 2018). The Phytopathology Laboratory has the accreditation for bacteria and viruses identification, so it seeks accreditation to guarantee technical competence in its fungal tests using the BIOLOG microbial identification system, which is based on the identification of microorganisms through their metabolic profile using different carbon sources. The general objective of this research was to validate the analytical method for the diagnosis of Botrytis cinerea through the MICROBIAL IDENTIFICATION SYSTEM BIOLOG and ensure the quality of the assay through genetic analysis. For this validation, the ISO/IEC 17025 Standard criteria and the general accreditation criteria of the SAE were applied. A search through the laboratory database was initialy carried out in order to determine the presence and persistence of the pathogen in different provinces of the country. An isolation, identification and preservation of Botrytis sp. strains which affect several types of crop was necessary. This was achieved through the application of morphological, morphometric, biochemical, immunoassay and molecular techniques. Finally, the validation of the method and the assurance of the quality of the test was obtained, identifying the limitations and parameters that may affect the analyte. This research concluded with the positive identification of the pathogen in all the assays carried out mainly in the MICROBIAL IDENTIFICATION SYSTEM BIOLOG