The use of peroxidase anti-peroxidase (PAP) complexes in the detection of plant viruses by ELISA.
An ELISA test system for detection of plant viruses in field samples is described, based on the unlabelled antibody method using a peroxidase-antiperoxidase (PAP) complex. Novel features of the system include the use of acid-treated naked bacteria as combined carrier-adjuvants for the production of rabbit antiviral antibodies, and the use of acid-treated chicken antibodies (IgY) for antigen trapping in the ELISA. Systems for detection of potato virus Y (PVY), potato leafroll virus (PLRV), grapevine fanleaf virus (GFV) and grapevine virus A (GVA) were developed and compared with conventional direct double antibody sandwich (DAS) systems in tests with both purified virus and field samples. The PAP systems offer improved sensitivity, no background problems in the outer rows of the microtitre plates and are much easier to visualize with the naked eye if no plate reader is available.
Rowland
G F
GF
Bioclones (Pty) Ltd, c/o Department of Biochemistry, University of Stellenbosch, Republic of South Africa.
Engelbrecht
D J
DJ
Pool
E J
EJ
Schmollgruber
E C
EC
Thompson
G J
GJ
van der Merwe
K J
KJ
D000818:Animals; D000914:Antibodies, Viral / Q000096:biosynthesis; D000956:Antigens, Viral / Q000276:immunology; D002645:Chickens; D004797:Enzyme-Linked Immunosorbent Assay; D006041:Goats; D007124:Immunoenzyme Techniques; D010942:Plant Viruses / Q000276:immunology / Q000302:isolation & purification*; D011817:Rabbits; D011933:Reagent Kits, Diagnostic
D000914:Antibodies, Viral; D000956:Antigens, Viral; D011933:Reagent Kits, Diagnostic