T7 phage display of Ep15 peptide for the detection of WNV IgG.
West Nile virus (WNV) is one of the major emerging infectious diseases in North America. WNV belongs to the genus Flavivirus, and its rapid and extensive global spread has highlighted the necessity for accurate and specific assays for diagnosis of WNV infection. This study presents the first phage displayed peptide based ELISA for detection of WNV immunoglobulin G (IgG). The Ep15 epitope, derived from the WNV E protein DIII, was cloned into a T7 phage display system that was then used as recombinant antigen in a chemiluminescent ELISA format. The phage concentration was optimized at 5 x 10(10)PFU/ml and was used directly after polyethylene glycol concentration. The assay shows a limit of detection at a serum titer of 1:51,200 and a dynamic range from 1:100 to 1:2000. A screen of a panel of 66 human sera samples, and comparison with a commercial kit, revealed a sensitivity of 67% and a specificity of 100%. Considering the ease of antigen preparation, its stability and the optimum display properties of the T7 bacteriophage, it is apparent that this approach can be useful for the preparation of highly sensitive and specific anti-WNV immunoglobulin diagnostic kits.
Herrmann
Sebastien
S
Department of Biotechnology Engineering, Ben-Gurion University of the Negev, Beer-Sheva 84105, Israel.
Leshem
Boaz
B
Lobel
Leslie
L
Bin
Hannah
H
Mendelson
Ella
E
Ben-Nathan
David
D
Dussart
Philippe
P
Porgador
Angel
A
Rager-Zisman
Bracha
B
Marks
Robert S
RS
D000914:Antibodies, Viral / Q000097:blood*; D017123:Bacteriophage T7 / Q000302:isolation & purification / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D000939:Epitopes / Q000378:metabolism; D006801:Humans; D007074:Immunoglobulin G / Q000097:blood*; D019151:Peptide Library; D011994:Recombinant Proteins / Q000378:metabolism; D012680:Sensitivity and Specificity; D014764:Viral Proteins / Q000276:immunology; D014901:West Nile Fever / Q000097:blood / Q000175:diagnosis; D014902:West Nile virus / Q000276:immunology*
D000914:Antibodies, Viral; D000939:Epitopes; D007074:Immunoglobulin G; D019151:Peptide Library; D011994:Recombinant Proteins; D014764:Viral Proteins