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@article{Bailon:2023:10.12688/wellcomeopenres.18827.1,
author = {Bailon, N and Ramos, E and Alvarado, K and Bernaola, L and Wilson, J and Montoya, R and Valencia, T and Evans, CA and Datta, S},
doi = {10.12688/wellcomeopenres.18827.1},
journal = {Wellcome open research},
title = {A controlled evaluation of filter paper use during staining of sputum smears for tuberculosis microscopy.},
url = {http://dx.doi.org/10.12688/wellcomeopenres.18827.1},
volume = {8},
year = {2023}
}

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TY  - JOUR
AB  - <b>Background</b>: Some sputum smear microscopy protocols recommend placing filter paper over sputum smears during staining for <i>Mycobacterium tuberculosis</i> (TB) <i>.</i> We found no published evidence assessing whether this is beneficial. We aimed to evaluate the effect of filter paper on sputum smear microscopy results. <b>Methods:</b> 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.   <b>Results:</b> 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
AU  - Bailon,N
AU  - Ramos,E
AU  - Alvarado,K
AU  - Bernaola,L
AU  - Wilson,J
AU  - Montoya,R
AU  - Valencia,T
AU  - Evans,CA
AU  - Datta,S
DO  - 10.12688/wellcomeopenres.18827.1
PY  - 2023///
SN  - 2398-502X
TI  - A controlled evaluation of filter paper use during staining of sputum smears for tuberculosis microscopy.
T2  - Wellcome open research
UR  - http://dx.doi.org/10.12688/wellcomeopenres.18827.1
VL  - 8
ER  -

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