Evaluation of Trypan Blue Stain in a Haemocytometer for Rapid Detection of Cerebrospinal Fluid Sterility in HIV Patients with Cryptococcal Meningitis

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dc.contributor.authorKwizera, Richard
dc.contributor.authorAkampurira, Andrew
dc.contributor.authorKandole, Tadeo K.
dc.contributor.authorKambugu, Andrew
dc.contributor.authorKambugu, Andrew
dc.contributor.authorMeya, David B.
dc.contributor.authorBoulware, David R.
dc.contributor.authorRhein, Joshua
dc.contributor.authoron behalf of the ASTRO-CM Study Team
dc.date.accessioned2022-08-19T14:46:41Z
dc.date.available2022-08-19T14:46:41Z
dc.date.issued2017
dc.description.abstractQuantitative culture is the most common method to determine the fungal burden and sterility of cerebrospinal fluid (CSF) among persons with cryptococcal meningitis. A major drawback of cultures is a long turnaround-time. Recent evidence demonstrates that live and dead Cryptococcus yeasts can be distinguished using trypan blue staining. We hypothesized that trypan blue staining combined with haemocytometer counting may provide a rapid estimation of quantitative culture count and detection of CSF sterility. To test this, we evaluated 194 CSF specimens from 96 HIV-infected participants with cryptococcal meningitis in Kampala, Uganda. Cryptococcal meningitis was diagnosed by CSF cryptococcal antigen (CRAG). We stained CSF with trypan blue and quantified yeasts using a haemocytometer. We compared the haemocytometer readings versus quantitative Cryptococcus CSF cultures. Haemocytometer counting with trypan blue staining had a sensitivity of 98% (64/65), while CSF cultures had a sensitivity of 95% (62/65) with reference to CSF CRAG for diagnostic CSF specimens. For samples that were positive in both tests, the haemocytometer had higher readings compared to culture. For diagnostic specimens, the median of log10 transformed counts were 5.59 (n = 64, IQR = 5.09 to 6.05) for haemocytometer and 4.98 (n = 62, IQR = 3.75 to 5.79) for culture; while the overall median counts were 5.35 (n = 189, IQR = 4.78–5.84) for haemocytometer and 3.99 (n = 151, IQR = 2.59–5.14) for cultures. The percentage agreement with culture sterility was 2.4% (1/42). Counts among non-sterile follow-up specimens had a median of 5.38 (n = 86, IQR = 4.74 to 6.03) for haemocytometer and 2.89 (n = 89, IQR = 2.11 to 4.38) for culture. At diagnosis, CSF quantitative cultures correlated with haemocytometer counts (R2 = 0.59, P < 0.001). At 7–14 days, quantitative cultures did not correlate with haemocytometer counts (R2 = 0.43, P = 0.4). Despite a positive correlation, the haemocytometer counts with trypan blue staining did not predict the outcome of quantitative cultures in patients receiving antifungal therapy.en_US
dc.identifier.citationKwizera, R., Akampurira, A., Kandole, T. K., Nielsen, K., Kambugu, A., Meya, D. B., ... & Rhein, J. (2017). Evaluation of trypan blue stain in a haemocytometer for rapid detection of cerebrospinal fluid sterility in HIV patients with cryptococcal meningitis. BMC microbiology, 17(1), 1-7.https://doi.org/10.1186/s12866-017-1093-4en_US
dc.identifier.issn1471-2180
dc.identifier.urihttps://nru.uncst.go.ug/handle/123456789/4327
dc.language.isoenen_US
dc.publisherBMC microbiologyen_US
dc.subjectTrypan blue, Cryptococcal meningitis, HIV, Diagnostic techniques and procedures, Point-of-care systemsen_US
dc.titleEvaluation of Trypan Blue Stain in a Haemocytometer for Rapid Detection of Cerebrospinal Fluid Sterility in HIV Patients with Cryptococcal Meningitisen_US
dc.typeArticleen_US
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