Development and evaluation of a rapid immunomagnetic bead assay for the detection of classical swine fever virus antigen.
Classical swine fever (CSF) is a highly contagious and severe viral disease of swine resulting in substantial production losses in different farming systems in many regions of the world. The accurate and rapid detection of CSF outbreaks is reliant on sensitive and specific laboratory testing and is a key component of disease control. Specific detection of CSF virus can be achieved by virus isolation in tissue culture, antigen capture or the detection of viral RNA using molecular techniques. In order to reduce the time taken to achieve a diagnostic result and simplify testing methods, an antigen capture ELISA using immunomagnetic beads (IMB) as the solid phase was developed and compared to a microplate-based antigen capture (AC)-ELISA. The IMB-ELISA has up to 64-fold greater analytical sensitivity than the AC-ELISA and initial estimates of diagnostic sensitivity and specificity are 100%. The IMB-ELISA has a highly robust, rapid and stable test format and is simpler to perform than the AC-ELISA. The IMB-ELISA has the added advantage that a result can be sensitively and specifically determined by eye, lending it to the possibility of adaptation to a near-to-field test with minimal equipment or expertise needed.
Conlan
James V
JV
CSIRO Australian Animal Health Laboratory, Geelong, Victoria, Australia. jvconlan@hotmail.com
Khounsy
Syseng
S
Blacksell
Stuart D
SD
Morrissy
Christopher J
CJ
Wilks
Colin R
CR
Gleeson
Laurence J
LJ
D000818:Animals; D000956:Antigens, Viral / Q000032:analysis*; D006692:Classical Swine Fever Virus / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary; D018189:Immunomagnetic Separation / Q000379:methods / Q000662:veterinary*; D013552:Swine; D013553:Swine Diseases / Q000175:diagnosis* / Q000276:immunology / Q000821:virology*
D000956:Antigens, Viral