A simple sandwich ELISA (WELYSSA) for the detection of lyssavirus nucleocapsid in rabies suspected specimens using mouse monoclonal antibodies.
Monoclonal antibody (MAb)-based capture enzyme-linked immunosorbent assays (ELISA) were developed for the diagnosis of rabies-suspect specimens. A combination of four mouse monoclonal antibodies directed against the rabies virus nucleocapsid was selected and used for the detection. The test was optimized and standardized so that maximum concordance could be maintained with the standard procedures of rabies diagnosis recommended by the WHO expert committee. Using prototype viruses from the different genotypes of lyssavirus and from various geographic origins and phylogenetic lineages, this paper presents a reliable, rapid and transferable diagnostic method, named WELYSSA that readily permits the detection of lyssaviruses belonging to the 7 genotypes of lyssavirus circulating in Europe, Africa, Asia and Oceania. The threshold of detection of lyssavirus nucleocapsids is low (0.8 ng/ml). With a panel of 1030 specimens received for rabies diagnostic testing, this test was found to be highly specific (0.999) and sensitive (0.970) when compared to other recommended rabies diagnostic methods.
Xu
Gelin
G
Wuhan Institute of Biological Products, Wuhan 430060, Hubei Province, China. xugelin@yahoo.com.cn
Weber
Patrick
P
Hu
Qiaoling
Q
Xue
Honggang
H
Audry
Laurent
L
Li
Chengping
C
Wu
Jie
J
Bourhy
Herve
H
D000818:Animals; D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D001921:Brain / Q000821:virology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods* / Q000706:statistics & numerical data; D005838:Genotype; D006801:Humans; D018114:Lyssavirus / Q000145:classification / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D051379:Mice; D019251:Nucleocapsid / Q000032:analysis* / Q000276:immunology; D011818:Rabies / Q000175:diagnosis* / Q000276:immunology / Q000821:virology; D011820:Rabies virus / Q000145:classification / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D018353:Rhabdoviridae Infections / Q000175:diagnosis / Q000276:immunology / Q000821:virology; D012680:Sensitivity and Specificity
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral