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A controlled evaluation of filter paper use during staining of sputum smears for tuberculosis microscopy.

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dc.contributor.author Bailon Gonzales, Nataly Aracely
dc.contributor.author Ramos Maguiña, Eric Sreve
dc.contributor.author Alvarado Torres, Keren Soledad
dc.contributor.author Bernaola Silva, Lenin Ceferino
dc.contributor.author Wilson, James
dc.contributor.author Montoya, Rosario
dc.contributor.author Valencia, Teresa
dc.contributor.author Evans, Carlton Anthony William
dc.contributor.author Datta, Sumona
dc.date.accessioned 2023-10-12T15:30:07Z
dc.date.available 2023-10-12T15:30:07Z
dc.date.issued 2023
dc.identifier.uri https://hdl.handle.net/20.500.12866/14278
dc.description.abstract Background: Some sputum smear microscopy protocols recommend placing filter paper over sputum smears during staining for Mycobacterium tuberculosis (TB) . We found no published evidence assessing whether this is beneficial. We aimed to evaluate the effect of filter paper on sputum smear microscopy results. Methods: Sputum samples were collected from 30 patients with confirmed pulmonary TB and 4 healthy control participants. From each sputum sample, six smears (204 smears in total) were prepared for staining with Ziehl-Neelsen (ZN), auramine or viability staining with fluorescein diacetate (FDA). Half of the slides subjected to each staining protocol were randomly selected to have Whatman grade 3 filter paper placed over the dried smears prior to stain application and removed prior to stain washing. The counts of acid-fast bacilli (AFB) and precipitates per 100 high-power microscopy fields of view, and the proportion of smear that appeared to have been washed away were recorded. Statistical analysis used a linear regression model adjusted by staining technique with a random effects term to correct for between-sample variability.   Results: The inclusion of filter paper in the staining protocol significantly decreased microscopy positivity independent of staining with ZN, auramine or FDA (p=0.01). Consistent with this finding, there were lower smear grades in slides stained using filter paper versus without (p=0.04), and filter paper use reduced AFB counts by 0.28 logarithms (95% confidence intervals, CI=0.018, 0.54, p=0.04) independent of staining technique. In all analyses, auramine was consistently more sensitive with higher AFB counts versus ZN (p=0.001), whereas FDA had lower sensitivity and lower AFB counts (p<0.0001). Filter paper use was not associated with the presence of any precipitate (p=0.5) or the probability of any smear washing away (p=0.6) during the staining process. Conclusions: Filter paper reduced the sensitivity of AFB microscopy and had no detectable beneficial effects so is not recommended. en_US
dc.language.iso eng
dc.publisher F1000 Research
dc.relation.ispartofseries Wellcome Open Research
dc.rights info:eu-repo/semantics/restrictedAccess
dc.rights.uri https://creativecommons.org/licenses/by-nc-nd/4.0/deed.es
dc.subject Filter paper en_US
dc.subject Tuberculosis en_US
dc.subject Ziehl Neelsen en_US
dc.subject Auramine en_US
dc.subject Fluorescein diacetate en_US
dc.subject Sputum smear en_US
dc.subject Acid-fast bacilli en_US
dc.subject Microscopy en_US
dc.subject.mesh Papel
dc.subject.mesh Tuberculosis
dc.subject.mesh Benzofenoneido
dc.subject.mesh Colorantes Fluorescentes
dc.subject.mesh Microscopía
dc.subject.mesh Enterobacteriaceae
dc.title A controlled evaluation of filter paper use during staining of sputum smears for tuberculosis microscopy. en_US
dc.type info:eu-repo/semantics/article
dc.identifier.doi https://doi.org/10.12688/wellcomeopenres.18827.1
dc.relation.issn 2398-502X


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