Abstract
Purpose: An unprecedented Ebola outbreak occurred in 2014-2015 in West Africa. A better understanding of the EBOV life cycle is fundamental to develop new countermeasures, as well as to fully comprehend the pathways of inter-human transmission. We have explored the possibility of viral persistence in different body fluids’ samples obtained from a Health Care Worker (HCW) infected in Sierra Leone and treated at INMI L. Spallanzani, Italy. To evaluate whether the virus was in a replicative status or simply derived from blood spill over, we compared the trends of EBOV-specific negative sense genomic RNA (neg-RNA), positive sense RNA (pos-RNA) and total viral RNA in different clinical samples. Methods & Materials: Clinical samples were inactivated in BSL4 facility and RNA was extracted with QIAamp Viral-RNA Mini-Kit (Qiagen). Quantification of total viral RNA was performed with the reference test Altona-FilovirusScreen Kit1.0 (Altona Diagnostics). To measure EBOV-specific negative sense genomic RNA (neg-RNA) and positive sense RNA (pos-RNA) [including both replication intermediate (cRNA) and messenger RNA (mRNA)], L-gene specific reverse or forward primers were used in the reverse transcription step Results: The clinical samples analyzed (sputum, nasopharyngeal swab, ocular swab, urine and plasma) show different trends in neg-RNA, pos-RNA and total viral RNA levels. In Nasopharyngeal swab, Ocular Swab and Urine, total viral RNA is the only one detectable, until Day 12, 5 and 15 after hospitalization, respectively. On the contrary, in plasma, total viral RNA, neg-RNA and pos-RNA levels simultaneously decreased, starting from day 3 and becoming undetectable at Day 6. In Sputum, pos-RNA levels decreased since Day 8, persisting at detectable levels up to Day 10, coherently with total viral RNA and neg-RNA levels, which start decreasing at Day 10 and become undetectable at Day 11. Conclusion: The presence of viral RNA and replication markers (pos-RNA and neg-RNA) in sputum might be of relevance in future analysis regarding the persistence of the virus in the upper respiratory tract, even after viral clearance from plasma. These results should be taken under further investigation in order to better understand the role of the respiratory tract for possible involvement in viral shedding, viral replication site or as a viral reservoir.
Highlights
40 of 54 finished questionnaires were analysed
Most local public health departments in North Rhine-Westphalia (NRW) had to deal with the Ebola topic, but 23 of the participants did not report any probable Ebola case and 29 less than 5 suspected cases
82% of participants think to be better prepared for suspected cases of highly pathogenic infectious diseases
Summary
40 of 54 finished questionnaires were analysed. Of those 11 were completed in an urban district and 26 in a rural district in NRW (3 not stated). Most local public health departments in NRW had to deal with the Ebola topic, but 23 of the participants did not report any probable Ebola case and 29 less than 5 suspected cases. 26 participants stated to have purchased PPE in their public health office, only 19 have trained donning and doffing of PPE and mere 5 are planning to continue PPE training. 82% of participants think to be better prepared for suspected cases of highly pathogenic infectious diseases. Half the participants used the epidemics preparedness plan NRW and found it to be helpful, but 25% wished to receive relevant information sooner
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have