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Detection of Abroviruses and Arbovirus Infections Diagnosis

#0624


Detection of Abroviruses and Abrovirus Infections Diagnosis by Time-Resolved Fluoroimmunoassay

Tech Area / Field

  • BIO-CGM/Cytology, Genetics and Molecular Biology/Biotechnology
  • BIO-CHM/Biochemistry/Biotechnology
  • BIO-OTH/Other/Biotechnology

Status
8 Project completed

Registration date
16.04.1996

Completion date
30.09.2005

Senior Project Manager
Weaver L M

Leading Institute
State Research Institute of Biological Instrument-Making, Russia, Moscow

Collaborators

  • Institut Pasteur, France, Paris\nUniversity of London / London School of Hygiene&Tropical Medicine, UK, London\nUniversity of Texas / Medical Branch, USA, TX, Galveston

Project summary

Aim of this project is the development of time-resolved fluoroimmunoassay (TR-FIA) method for diagnosis of actual for Russia natural-foci arbovirus infections (Tick-borne encephalitis, Crimean hemorrhagic fever, Heirorrhagic fever with renal syndrome and others) and detection of these viruses in natural, clinical and experimental samples.

Detection of specific virus antigens, RNA, IgG and IgM antibodies will be carried out on the basis of common approach - time-resolved fluoroimmunoassay. This technology is based on the use of lanthanide chelate labels with unique fluorescence properties, time-resolved fluorometry and dissociation-enhancement principle. TR-FIA is a method of choice among other non-isotopic methods (conventional fluorescence, enzyme immunoassay) as well as radioimmunoassay due to high sensitivity, wide dynamic range, stable fluorescence and labeled compounds, reproducible results, possibility of dual, triple and quadruple labeling.

Direction of research


1. Use of new luminescent labels of metalloporphyrine group and rare-earth metal group for simultaneous detection of different antigens in one sample.
2. Detection of viruses in clinical samples and vectors (ticks and mosquitoes).
3. Determination of cut-off levels of virus detection taking into account the concentration of specific virus proteins, RNA and infectious activity of viruses.
4. Use of obtained data for seroprophylactics of tick-borne encephalitis in persons which were bitten by infected ticks.
5. Determination of IgM and IgG specific antibodies in patients and for seroepidemiological investigations.
6. Study of antigenic structure and classification of viruses and development of diagnostic TR-FIA test-systems.

In the base of the project are the works of GosNII-BP and the results of the preceeding research made in co-operation with Ivanovsky Institute of Virology RAMS, Moscow, Russia.

Expected results

Theoretical meaning

New data about antigenic structure of examined viruses allows to extend on evolution of arboviruses, improve their classification and develop the new approaches to detection and identification.

Practical meaning


- Recommendations on seroprophilactics of TBE in persons bitten by infected ticks will be developed.
- Complex of methods based on common approach for reliable detection and identification of viruses will be proposed.
- Information and normative documents for production of TR-FIA test-systems will be developed.
- TR-FIA test-systems will be used for early as well as retrospective diagnosis of arbovirus infections and seroepidemiological studies.

We welcome colleagues from Europe, USA and Japan for participation in project fulfillment in any suitable form.

Participation of foreign collaborators is desirable in following points:


1. Cooperative analysis and evaluation of results of investigation.
2. Exchange with scientific information.
3. Exchange with bioreagents (probes, immune sera, monoclonal antibodies, strains of viruses, diagnostic samples) useful for the project fulfillment.


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