Digital rolling circle amplification-based detection of Ebola and other tropical viruses

Publication date: Available online 16 December 2019Source: The Journal of Molecular DiagnosticsAuthor(s): Sibel Ciftci, Felix Neumann, Samir Abdurahman, K. Sofia Appelberg, Ali Mirazimi, Mats Nilsson, Narayanan MadaboosiAbstractEmerging tropical viruses have caused serious outbreaks during the recent years, such as Ebola virus (EBOV) in 2014 and the most recent in 2018-19 in Congo. Thus, immediate diagnostic attention is demanded at the point-of-care in resource-limited settings, since the performance and the operational parameters of conventional EBOV testing are limited. Especially, their sensitivity, specificity and coverage of other tropical disease viruses make them unsuitable for diagnostic at the point-of-care. Here, we present a padlock probe (PLP)-based rolling circle amplification (RCA) method for the detection of EBOV. For this, a set of PLPs, separately targeting the viral RNA (vRNA) and complementary RNA (cRNA) of all the seven genes of EBOV, was used in the RCA assay and validated on virus isolates from cell culture. The assay was then translated for testing clinical samples, and simultaneous detection of both EBOV RNA types was demonstrated. For increased sensitivity, the RCA products were enriched on a simple and pump-free microfluidic chip. As PLPs and RCA are inherently multiplexable, we demonstrate the extension of the probe panel for the simultaneous detection of the tropical viruses Ebola, Zika and Dengue. The demonstrated high specificity, sensitivity an...
Source: The Journal of Molecular Diagnostics - Category: Pathology Source Type: research

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Source: The Journal of Molecular Diagnostics - Category: Pathology Source Type: research
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This report summarizes the lectures held at the meeting and highlights advances in the field. PMID: 30597184 [PubMed - as supplied by publisher]
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More News: Congo Health | Dengue Fever | Ebola | Genetics | Outbreaks | Pathology | Zika Virus