Abstract
The aim of our study was to find out the optimal conditions for short-term storage of cerebrospinal fluid (CSF) samples for direct diagnosis of Lyme disease. A mixture of Borrelia-negative CSFs spiked with a defined amount of cultured Borrelia garinii was used. Borrelia stability was investigated over 7 days at four different temperatures [room temperature (RT), +4, −20 and −70 °C]. Quantitative changes in CSF Borrelia were measured by quantitative PCR (qPCR), and morphological changes in the spirochetes were observed by transmission electron microscopy (TEM). These qPCR results were statistically evaluated. We found +4 °C to be an optimal temperature for short-term storage of CSF samples intended for TEM observation. There was no significant difference between the temperatures tested in the average quantity of Borrelia measured by qPCR. On the contrary, electron optical diagnosis of frozen samples and samples stored at RT showed destructive morphological changes and decreased spirochete counts. Our results show that optimal conditions for the pre-analytical phase of investigation of one type of material can differ depending on the diagnostic method employed.
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Acknowledgments
This study was supported by the Grant Agency of the Ministry of Health of the Czech Republic (IGA MZ ČR—No. NT/13467 - 4) and by MH CZ - DRO (The National Institute of Public Health—NIPH, 75010330).
Compliance with ethical standards
The authors assert that all procedures contributing to this work comply with the ethical standards of the relevant national and institutional committees on human experimentation and with the Helsinki Declaration of 1975, as revised in 2008.
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Berenová, D., Krsek, D., Šípková, L. et al. Short-term stability of Borrelia garinii in cerebrospinal fluid. Folia Microbiol 61, 45–50 (2016). https://doi.org/10.1007/s12223-015-0408-9
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DOI: https://doi.org/10.1007/s12223-015-0408-9