Etablierung und Evaluation eines vereinfachten, speziesunabhängigen Anti-RVFV-Nukleoprotein-ELISAs

Establishment and evaluation of a simplified, species-independent anti-RVFV-nucleoprotein ELISA

by Uurtsaikh Schmidt née Balchin
Doctoral thesis
Date of Examination:2021-12-08
Date of issue:2021-12-07
Advisor:Prof. Dr. Frank T. Hufert
Referee:Prof. Dr. Frank T. Hufert
Referee:Prof. Dr. Stefan Pöhlmann
Persistent Address: http://dx.doi.org/10.53846/goediss-8994

 

 

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Abstract

English

The Rift-Valley-fever-virus (RVFV) belongs to the family Phenuiviridae and is transmitted by insect bite. It is the causative agent of Rift Valley fever disease which affects livestock and humans in Africa and the Arabian Peninsula. Competitive as well as indirect ELISA methods that detect antibodies against the RVFV-nucleoprotein (RVFV-N) are a routinely available screening method even in poorly equipped laboratories. The competitive ELISA is species independent but not very sensitive while the more sensitive indirect ELISA depends on species-specific secondary antibodies. In this thesis, a new ELISA method has been developed that combines species-independence with good sensitivity and specificity. To this aim, recombinantly expressed RVFV-N was coupled to horseradish peroxidase to detect serum antibodies that are bound to RVFV-N coated wells. This newly established ELISA could detect RVFV-N antibodies in 1 µl serum samples from different species with a sensitivity and specificity comparable to the competitive ELISA, which requires 50 µl serum for each test. Additionally, a bacterial expression system for the RVFV-Gn surface protein was established. The recombinant RVFV-Gn protein can be used to develop a species-independent for the detection of neutralizing RFVF-Gn antibodies. The combination of ELISA tests detecting RVFV-N antibodies and RVFV-G antibodies may also help to discriminate between infected and vaccinated animals.
Keywords: ELISA; RVFV
Schlagwörter: ELISA; RVFV
 
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