title: Short-term stability of Borrelia garinii in cerebrospinal fluid.
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.
authors.affiliation: National Reference Laboratory for Lyme Borreliosis, The National Institute of Public Health, Šrobárova 48, 100 42, Prague 10, Czech Republic. berenova@seznam.cz.
authors.forename: Dagmar
authors.initials: D
authors.lastname: Berenová
authors.affiliation: National Reference Laboratory for Lyme Borreliosis, The National Institute of Public Health, Šrobárova 48, 100 42, Prague 10, Czech Republic.
authors.forename: Daniel
authors.initials: D
authors.lastname: Krsek
authors.affiliation: National Reference Laboratory for Lyme Borreliosis, The National Institute of Public Health, Šrobárova 48, 100 42, Prague 10, Czech Republic.
authors.forename: Lenka
authors.initials: L
authors.lastname: Šípková
authors.affiliation: National Reference Laboratory for Lyme Borreliosis, The National Institute of Public Health, Šrobárova 48, 100 42, Prague 10, Czech Republic.
authors.forename: Alena
authors.initials: A
authors.lastname: Lukavská
authors.affiliation: Department of Biostatistics and Informatics, The National Institute of Public Health, Šrobárova 48, 100 42, Prague 10, Czech Republic.
authors.forename: Marek
authors.initials: M
authors.lastname: Malý
authors.affiliation: National Reference Laboratory for Lyme Borreliosis, The National Institute of Public Health, Šrobárova 48, 100 42, Prague 10, Czech Republic.
authors.forename: Zuzana
authors.initials: Z
authors.lastname: Kurzová
authors.affiliation: National Reference Laboratory for Lyme Borreliosis, The National Institute of Public Health, Šrobárova 48, 100 42, Prague 10, Czech Republic.
authors.forename: Jan
authors.initials: J
authors.lastname: Hořejší
authors.affiliation: National Reference Laboratory for Lyme Borreliosis, The National Institute of Public Health, Šrobárova 48, 100 42, Prague 10, Czech Republic.
authors.forename: Petr
authors.initials: P
authors.lastname: Kodym
mesh_terms: D015748:Borrelia burgdorferi Group / Q000502:physiology*; D002555:Cerebrospinal Fluid / Q000382:microbiology*; D006801:Humans; D008193:Lyme Disease / Q000175:diagnosis* / Q000382:microbiology; D050296:Microbial Viability; D013048:Specimen Handling / Q000379:methods*; D013696:Temperature; D013997:Time Factors