Abstract
In 2013–2016, West Africa experienced the largest and longest Ebola virus disease outbreak ever documented. The wide geographic spread and magnitude of the outbreak often limited the timely and rapid testing of diagnostic samples from patients with suspected Ebola virus disease, raising questions regarding the optimal storage and shipping conditions of clinically relevant specimens, including EDTA-whole blood, plasma, capillary blood, urine and seminal fluid (associated with sexual transmission of the Ebola virus after recovery from the disease). Therefore, the aim of our study was to identify the extent to which storage temperature and clinical specimen type influence Ebola virus viability. Virus infectivity was determined using a fluorescent focus-forming assay. In our study, we show that Ebola virus was the most stable in EDTA-whole blood and plasma samples, whereas rapid decay of infectivity was observed in simulated capillary blood, urine and semen samples, especially when these samples were stored at higher temperatures. The analysis of variance results demonstrated that both temperature and clinical specimen type have significant effects on virus viability, whereas donor differences were not observed. Repeated freeze and thaw cycles of the samples also had a notable impact on virus viability in EDTA-whole blood and urine. Due to the rapid temperature- and specimen-dependent degradation of the virus observed here, our study highlights the importance of proper clinical sample storage at low temperatures during transportation and laboratory analysis.
Highlights
Ebola outbreaks, especially those that occurred in2013–2016 in West Africa, pose significant challenges for outbreak response teams with respect to ensuring adequate conditions for the collection, transport, storage and processing of different biological specimens for laboratory testing[1]
The samples, which were obtained from healthy volunteers, were spiked with Ebola virus (EBOV) Makona strain to achieve a final titer of 105 fluorescent focus unit (FFU)/mL; the samples were stored at 37, 23 and 4 ° C and at −80 °C as a control
Virus viability was most stable in EDTAwhole blood (Fig. 1a) and plasma (Fig. 1b) with titer reductions of 1.56 and 1.42 log[10], respectively, at 37 °C after 120 h
Summary
2013–2016 in West Africa, pose significant challenges for outbreak response teams with respect to ensuring adequate conditions for the collection, transport, storage and processing of different biological specimens for laboratory testing[1]. Shipping of the samples to reference laboratories during the first 48 h after collection is highly recommended and may influence test reliability[2]. The RNA of Ebola virus (EBOV) is known to be relatively stable in whole blood and serum/plasma samples for weeks under African environmental conditions, depending on the initial viral load[3,4,5]. The environmental stability of the virus has been studied, and the potential persistence of its viral RNA under hospital circumstances has been determined[9,10]. EBOV RNA in Ebola treatment centers was found to be associated with surfaces that come into close contact with patients and infectious body
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