Detection of dengue viral RNA by microplate hybridization.
Dengue virus infection is a major public health problem throughout tropical countries. In endemic areas, dengue hemorrhagic fever (DHF) or dengue shock syndrome (DSS) are common complications resulting in death. However, serological confirmation of dengue-related illness is often complicated and time-consuming. Detection of dengue viruses in clinical or field samples usually depends on virus isolation in susceptible cell lines or in mosquitoes, followed by viral protein identification using polyclonal or monoclonal antibodies. The increasing incidence of dengue virus infections has prompted increased efforts to develop rapid and reliable diagnostic techniques. A simple microplate hybridization method was developed for identification of viral RNA. Microplate hybridization is simpler than enzyme-linked immunosorbent assay and has several advantages over the conventional dot-blot hybridization method: (1) radioisotopes are not necessary; (2) synthetic oligonucleotide for the probe is not needed; (3) the time required for washing of the solid phase is greatly reduced; and (4) baking is eliminated. The results show that this procedure is sensitive, rapid and easy to perform.
Department of Molecular Biology, National University of Mexico, Ciudad Universitaria, México, D.F.
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Ruiz
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Zamora
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Liu
D000818:Animals; D001483:Base Sequence; D002522:Chlorocebus aethiops; D017931:DNA Primers; D015342:DNA Probes; D003715:Dengue / Q000097:blood / Q000175:diagnosis / Q000821:virology*; D003716:Dengue Virus / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction; D012367:RNA, Viral / Q000032:analysis*; D012194:RNA-Directed DNA Polymerase; D012680:Sensitivity and Specificity; D014709:Vero Cells
D017931:DNA Primers; D015342:DNA Probes; D012367:RNA, Viral; D012194:RNA-Directed DNA Polymerase