2. Aim of the project
• Aim: To develop a multiplex one-step reverse-transcription PCR
for use with a EuroArray for a range of emerging diseases and
their various heterotypic serotypes.
• An emerging disease is defined by the CDC as a disease in
which incidence in humans has increased in the past 20 years
and/or could increase in the future.
• Main task that I performed; Developing the “Small panel” test for;
Dengue Virus (1-4), Chikungunya Virus and Zika Virus (Africa and
Asia).
• "Large" panel currently consists of a further range of emerging
diseases including; CCHFV, YFV, RVFV and JEV.
3. Background into Diseases.
Problem with diagnosis of Dengue Virus, Zika Virus and Chikungunya Virus;
• Hard to differentiate between clinical manifestations
• Similar in their respective genomes.
All viruses in this are RNA viruses meaning that a positive patient sample will contain
RNA and therefore a RT-PCR is necessary.
RT-PCR means Reverse Transcription PCR which means that it contains a reverse
transcriptase enzyme and a reverse transcription step in the PCR cycle which allows for
the enzyme to transcribe the RNA into DNA that is then used in a normal PCR
4. Introduction to Invitro-Transcription
The pathogens on this array are all RNA viruses and therefore the research and
development of this project is necessary to be performed using RNA and we
produce our own RNA templates. This is done in a process called Invitro-
Transcription DNA Template (Plasmid)
RNA Template (Invitro-Transcription)
PCR using Primers with T7 promotor
Transcribed into RNA using T7 RNA Polymerase
DNA digestion to remove residual Template
dsDNA
7. Payment at Euroimmun
This is a payed laboratory based placement based in the north of Germany.
Near to a small city called Lübeck which is near to the second biggest city in
Germany- Hamburg.
This for me was a paid placement.
On top of the money that you get payed there are also several benefits which
include:
Free accommodation.
Lunch time meal Monday to Friday free
Erasmus Funding (not sure if still available.
Student finance.
