A robust lentiviral pseudotype neutralisation assay for in-field serosurveillance of rabies and lyssaviruses in Africa.
The inflexibility of existing serological techniques for detection of rabies in surveillance constrains the benefit to be gained from many current control strategies. We analysed 304 serum samples from Tanzanian dogs for the detection of rabies antibodies in a pseudotype assay using lentiviral vectors bearing the CVS-11 envelope glycoprotein. Compared with the widely used gold standard fluorescent antibody virus neutralisation assay, a specificity of 100% and sensitivity of 94.4% with a strong correlation of antibody titres (r=0.915) were observed with the pseudotype assay. To increase the assay's surveillance specificity in Africa we incorporated the envelope glycoprotein of local viruses, Lagos bat virus, Duvenhage virus or Mokola virus and also cloned the lacZ gene to provide a reporter element. Neutralisation assays using pseudotypes bearing these glycoproteins reveal that they provide a greater sensitivity compared to similar live virus assays and will therefore allow a more accurate determination of the distribution of these highly pathogenic infections and the threat they pose to human health. Importantly, the CVS-11 pseudotypes were highly stable during freeze-thaw cycles and storage at room temperature. These results suggest the proposed pseudotype assay is a suitable option for undertaking lyssavirus serosurveillance in areas most affected by these infections.
Wright
Edward
E
Division of Infection and Immunity, University College London, 46 Cleveland Street, London, United Kingdom. edward.wright@ucl.ac.uk
McNabb
Suzanne
S
Goddard
Trudy
T
Horton
Daniel L
DL
Lembo
Tiziana
T
Nel
Louis H
LH
Weiss
Robin A
RA
Cleaveland
Sarah
S
Fooks
Anthony R
AR
D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D004283:Dog Diseases / Q000175:diagnosis / Q000453:epidemiology / Q000821:virology; D004285:Dogs / Q000821:virology*; D005260:Female; D016086:Lentivirus / Q000235:genetics; D018114:Lyssavirus / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D008297:Male; D009500:Neutralization Tests; D011818:Rabies / Q000175:diagnosis / Q000453:epidemiology* / Q000662:veterinary; D011819:Rabies Vaccines / Q000008:administration & dosage; D011820:Rabies virus / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D012680:Sensitivity and Specificity; D016036:Seroepidemiologic Studies; D013636:Tanzania / Q000453:epidemiology; D014759:Viral Envelope Proteins / Q000235:genetics
D000914:Antibodies, Viral; D011819:Rabies Vaccines; D014759:Viral Envelope Proteins