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Title: LED fluorescence microscopy in the diagnosis of tuberculosis: Fading and restaining of smears for external quality assessment
Authors: Allassia, Sonia 
Aranibar, Mónica 
Boutonnet, Mónica 
Caserío, Viviana 
Etchart, Ana Alicia 
Fajardo, Sandra 
García, Mónica 
Gomez, Noemí 
Gunia, Alba Marisa 
Gustincic, María Virginia 
Izquierdo, Viviana 
Jara, Arnaldo Andrés 
Kozicky, Graciela 
Matteo, Mario 
Pellegrini, Carlos 
Pellegrino, Silvia 
Pérez Catalán, Sebastián 
Poggi, Susana 
Sacramone, Carina 
Santiso, Gabriela María 
Souto, Alejandro 
Togneri, Ana María 
Wolff, Lidia 
Vilche, Sandra 
Eletti, Daniel 
Imaz, María Susana 
Keywords: Microscopía de fluorescencia LED;Evaluación externa de calidad;Tuberculosis
Issue Date: 2016
Journal: Revista Argentina de microbiologia 
Blinded rechecking is a method proposed for external quality assurance (EQA) of auramine-stained acid-fast bacilli (AFB) smears using fluorescence microscopy (FM), however, this procedure is not well developed and slides fading over time could compromise its implementation. Since bleaching of fluorescent molecules involves temperature-dependent chemical reactions, it is likely that low temperatures could slow down this process. We stored auramine-stained slides under different environmental conditions, including -20°C, and examined them over time. The slides stored in all the environments faded. At -20°C, fading was not reduced in relation to room temperature. Restaining and re-examining smears after five months showed that the slides containing saliva and storage at -20°C were associated with failure in AFB reappearance. In conclusion, the practice of freezing slides until they are viewed should be discouraged as it has a negative effect on blinded rechecking by reducing reading concordance after restaining. Specimen quality should be considered when interpreting FM-EQA results.
ISSN: 0325-7541
DOI: 10.1016/j.ram.2016.03.006
Appears in Collections:Publicaciones INER

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