Rapid detection of tick-borne encephalitis virus sequences by cDNA amplification coupled to a simple DNA enzyme immunoassay.

BACKGROUND

The tick-borne encephalitis virus (TBEV) causes many cases of severe neurological disease in Europe and Asia. Rapid and virus specific laboratory tests are needed for epidemiological studies and for diagnosis of human diseases.

OBJECTIVES

A newly developed colorimetric method, DNA enzyme immunoassay (DEIA), was applied to the detection of TBEV-specific sequences.

STUDY

DESIGN

Reverse transcription and polymerase chain reaction (PRC) were used for g genomic amplification. The specificity of PCR products was confirmed using an internal biotinylated probe from the 5'-NCR immobilized on streptavidin coated microtiter wells. The hybridization event was revealed by monoclonal antibodies to double stranded DNA in a standard ELISA reaction.

RESULTS

A total of 23 TBEV strains among them prototype strains and isolates from Germany and other European countries was investigated. In all cases hybridization with the internal probe has occurred.

CONCLUSION

The DEIA described here is a simple and rapid method facilitating the handling of a large number of samples and therefore seems to be especially valuable to surveillance studies of endemic areas and for diagnosis of human diseases.