Isothermal amplification for rapid molecular diagnosis of exotic virus that cause hemorrhagic fever and other severe symptoms

Christopher Erik Jensen

Studenteropgave: Speciale

Abstrakt

New outbreaks of virus with disastrous outcome emerge from time to time like the 2013 Ebola virus outbreak. Diagnostic methods already exist for exotic viruses like Ebola, but they are time consuming, expensive and complicated and they need a lot of equipment to work, equipment not necessarily available at the hotspot of such outbreaks. Many people including healthcare workers died during the outbreak and better and safer diagnostics are needed to constrain such events to avoid worldwide crises.
A rather novel method called loop-mediated isothermal amplification (LAMP), facilitate the features necessary for a field test, as it is fast, cheap, requires a minimum of equipment and most of all it is safe to use with the right precautions. We wanted to investigate if this method was suitable as a diagnostic tool for exotic single stranded RNA viruses and if it was possible to ease the interpretation method for a naked eye evaluation. In addition, we wanted to demonstrate a safe and easy method for nucleic acid purification from clinical samples that could be performed in a point of care situation to avoid biosafety level 4 precautions.
We developed assays capable of detecting Ebola virus, Marburg virus and all Dengue virus serotypes, six times as fast as a conventional diagnostic PCR tool for exotic viruses. We show, that LAMP can ease diagnostics, enable safe point of care and help constrain future outbreaks faster.

UddannelserMolekylærbiologi, (Bachelor/kandidatuddannelse) KandidatMedicinalbiologi, (Bachelor/kandidatuddannelse) Kandidat
SprogEngelsk
Udgivelsesdato23 jun. 2017
Antal sider63
VejledereLotte Jelsbak