RESUMO
According to the data of the Federal Service on Customers' Rights Protection and Human Well-being Surveillance, the enzyme-linked immunosorbent assay (ELISA) detects the tick-borne encephalitis virus (TBEV) in ticks more often than the polymerase chain reaction (PCR). The goal of this work was to compare TBEV detection efficiency in the ixodid ticks of different species with the commercial kits based on ELISA and real-time PCR. Ticks of five species were parenterally infected with 2-6 IgPFU of the European or Siberian TBEV subtypes. We formed randomized and encoded series of infected and intact ticks of different species, and in "blind" experiment analyzed the ticks on the TBEV presence with the kits based on ELISA and real-time PCR. ELISA and real-time PCR effectiveness of the TBEV detection in ticks was not affected by gender, species of ticks or presence of blood meal. The kits based on ELISA were less sensitive than those based on real-time PCR. ELISA effectiveness depended on the TBEV subtype. The presence of the false positive reactions and sensitivity of ELISA were affected by the protocols of reaction. The problem of the different TBEV prevalence in the field-collected ticks obtained with various methods remains to be studied.
Assuntos
Antígenos Virais/imunologia , Vírus da Encefalite Transmitidos por Carrapatos/isolamento & purificação , Ensaio de Imunoadsorção Enzimática/normas , Ixodidae/virologia , RNA Viral/genética , Reação em Cadeia da Polimerase em Tempo Real/normas , Animais , Antígenos Virais/genética , Vírus da Encefalite Transmitidos por Carrapatos/genética , Vírus da Encefalite Transmitidos por Carrapatos/imunologia , Reações Falso-Positivas , Feminino , Masculino , Camundongos , Coelhos , Sensibilidade e Especificidade , Carga ViralRESUMO
The significant antigenic crossovers between West Nile virus (WNV) and tick-borne encephalitis virus (TBEV) make the immunological diagnosis of these diseases difficult. The avidicity index of virus-specific class G immunoglobulins (IgG) was used as a criterion for the differentiation of an immune response to WNV or TBEV in patients and convalescents. The panels of the sera sampled from patients with tick-borne encephalitis and convalescents in the Novosibirsk and Tomsk Regions and in the Primorye Territory and from those with West Nile fever and convalescents in the Volgograd Region. The determination of the avidicity index could establish that in the convalescents' sera, the avidicity index of virus-specific IgG was much higher than that in the patients' sera in the acute phase of infection. In relation to heteroantigen, the avidicity index and the positivity coefficient were substantially less than those in the reaction with homoantigen. The findings have indicated that the determination of the value of the avidicity index of virus-specific IgG and the positivity coefficient makes it possible to differentiate West Nile fever and tick-borne encephalitis with confidence on the basis of solid-phase enzyme immunoassay in determining virus-specific IgG in the sera of patients and convalescents in different regions of Russia.