A chemiluminescence detection method of hantaviral antigens in neutralisation assays and inhibitor studies.
A protocol for chemiluminescence detection of hantaviral antigens in infected cell foci is described. This focus detection is based on the conversion of a substrate into a luminescent product by peroxidase-antibody conjugates; the emitted light of infected cell foci can easily be recorded by autoradiography or video imaging providing a hard copy for documentation. The main advantage of this method as compared to conventional immunochemical staining is a higher detection sensitivity due to the inherent magnification effect of luminescence causing an obvious boost in focal image and intensity. This enables reduction of (i) incubation time of virus-infected cells and (ii) amount of needed antibody for antigen detection in foci. This method is applied to a chemiluminescence focus reduction assay for the serotyping of hantavirus-specific neutralising antibodies in infected persons and for the determination of activity of antiviral agents against hantavirus.
Heider
H
H
Institut für Virologie, Universitätsklinikum Charité, Humboldt-Universität, D-10098 Berlin, Germany.
Ziaja
B
B
Priemer
C
C
Lundkvist
A
A
Neyts
J
J
Krüger
D H
DH
Ulrich
R
R
D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology; D000956:Antigens, Viral / Q000032:analysis*; D000998:Antiviral Agents / Q000494:pharmacology*; D001345:Autoradiography; D002522:Chlorocebus aethiops; D006476:Orthohantavirus / Q000145:classification / Q000187:drug effects* / Q000276:immunology / Q000302:isolation & purification*; D018778:Hantavirus Infections / Q000821:virology; D006801:Humans; D008163:Luminescent Measurements; D009500:Neutralization Tests; D012254:Ribavirin / Q000494:pharmacology*; D012703:Serotyping / Q000379:methods; D014709:Vero Cells / Q000821:virology
D000914:Antibodies, Viral; D000956:Antigens, Viral; D000998:Antiviral Agents; D012254:Ribavirin