Development of Smear-Sputum Slides with Acid Fast Bacilli Positive Using Polyvinylidene Fluoride Nanofiber Filters
Keywords:
Sputum smear slide, Polyvinylidene fluoride nanofiber, Acid fast bacilliAbstract
Acid-fast staining of sputum is a simple and widely used method for diagnosis of pulmonary tuberculosis worldwide.
This method is high specificity but requires the expertise of the operator in analysis. Therefore, quality assurance of the testing method is essential, with sputum smear slides used as one of the quality control materials. This study aimed to develop a method for the preparing sputum smear slides using cultured acid fast bacilli (AFB) which were dispersed by filtration through a polyvinylidene fluoride (PVDF) nanofiber membrane coated with polyvinyl alcohol (PVA). Four types of the homemade nanofiber filters as 2Hz 8M, 2Hz 4M, 3Hz 8M and 3Hz 4M with a porosity of up to 99.98 percent and filter thicknesses of 158, 222, 48 and 81 micrometers, respectively were compared for filtration efficiency. When used to filter AFB, the 2Hz 4M PVDF membrane produced sputum smear slides with a high number of single AFB (averaging 4.0 ± 2.8 cells per oil power field) and small clusters of 2-5 AFB per cluster (averaging 4.67 ± 0.2 clusters per oil power field). The proportion of single AFB by the 2Hz 4M was higher than the number of AFB clumping in clusters compared to the use of other types of filters. In the preparation process of sputum smear slides, the concentration of AFB could be adjusted to give the result of AFB 3+, AFB2+, and AFB 1+ according to the World Health Organization's reporting criteria by diluting AFB at a 1:10 ratio for each reporting level. This prepared slides closely resembled the actual patient sputum, showing leukocytes and welled-dispersed single AFB under microscopic examination. This study is the first to apply nanofiber membrane filtration for dispersing AFB in sputum smear preparation. This method is simple, practical, and suitable for use in quality assurance of acid-fast staining in clinical laboratories as well as in training healthcare personnel.
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