Abstract
ObjectiveBacterial meningitis is a life threatening condition that requires prompt recognition and treatment. Currently, Gram stain is widely used for the microscopic detection of bacterial pathogens in cerebrospinal fluid (CSF). In Nepal, fluorescent microscopes have been installed in laboratories as a part of the National tuberculosis control program. However, information on the utility of the acridine orange (AO) stain for the direct detection of bacteria in CSF samples in Nepal is not available. Therefore, this study aims to compare Gram stain and AO stain for the rapid detection of bacterial pathogens in CSF of clinically suspected meningitis cases in Kathmandu, Nepal.ResultsBacterial pathogens were detected in 9.30% (36/387) by either of the three tests, 9.04% (35/387) by AO stain, 8.27% (32/387) by culture and 6.46% (25/387) by Gram’s stain. Considering culture as a gold standard, the sensitivity of AO stain was higher than Gram stain. The specificity of AO stain was 98.87%. Detection and differentiation of the bacteria was much clear in AO staining than Gram staining. AO is a better alternative to Gram stain in the rapid detection of bacterial pathogens in CSF in the setting where fluorescent microscope is available.
Highlights
Bacterial meningitis is a life threatening condition that requires prompt recognition and treatment [1]
Bacterial pathogens were detected in 9.30% (36/387) by either of the three tests, 9.04% (35/387) by acridine orange (AO) stain, 8.27% (32/387) by culture and 6.46% (25/387) by Gram’s stain
Considering culture as a gold standard, the sensitivity of AO stain was higher than Gram stain
Summary
Bacterial pathogens were detected in 9.30% (36/387) by either of the three tests, 9.04% (35/387) by AO staining, 8.27% (32/387) by culture and 6.46% (25/387) by Gram’s staining. 6.46% of the samples were positive for all the three tests i.e. Gram’s stain, acridine orange stain and culture. The sensitivity of AO stain was higher than Gram stain. Correlation between the Gram and AO staining of the same sample was good (Spearman = 0.833). The difference in positivity (7 specimen missed by Gram stain) between the two staining procedures was statistically significant (p value < 0.001). Of the 7 AO positive and Gram stain negative specimens, 6 were culture positive for bacterial pathogens (Table 2). Detection and differentiation of the bacteria was much clear in AO stained smear than Gram stained smear (Fig. 1). Excluding the cost of fluorescent microscope, both AO stain and Gram’s stain have similar price
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