Development and evaluation of a rapid immunomagnetic bead assay for the detection of classical swine fever virus antigen

Abstract

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.

Novel potential diagnostic test for Mycobacterium tuberculosis complex using combined immunomagnetic separation (IMS) and PCR-CTPP

“…Surprisingly, the stability of the antibodycoupled magnetic beads was still satisfactory even 1 year storage at 2-8°C. Similar results regarding stability for even over 10 weeks have previously been obtained when combined antibody-coated magnetic bead-ELISA and IMS-PCR were performed for the detection of classical swine fever virus (Conlan et al 2009) and hepatitis A (Jothikumar et al 1998) respectively.…”

“…Similar results regarding stability for even over 10 weeks have previously been obtained when combined antibody‐coated magnetic bead‐ELISA and IMS‐PCR were performed for the detection of classical swine fever virus (Conlan et al . ) and hepatitis A (Jothikumar et al . ) respectively.…”

Evaluation of an IgY-Based Immunomagnetic Enzyme-Linked Immunosorbent Assay System for Detection of Circulating Schistosoma japonicum Antigen in Serum Samples from Patients in China

“…40 A modification of magnetic particles is needed to provide high-binding capacity and dispersion stability in magnetic bead-based techniques, especially in diagnostic studies. 41 It is difficult to explain the positive reaction for the magnetic immunoassay found in egg-negative cases with or without a positive reaction in the IHA. Despite the IHA result, because it only represents antibody level, which cannot adequately overcome the presence of an immunologic scar, these positive results should be carefully identified.…”

A rapid and sensitive method based on magnetic beads for the detection of hepatitis B virus surface antigen in human serum

“…In contrast to physical adsorption, covalent bond formation between the antibody and magnetic beads leads to a much stronger association. Furthermore, antibody‐coated magnetic beads suspended in the reaction solution could increase the reaction rate and consequently reduce reaction times . All of these advantages result in an appreciable improvement in detection limit, reduction in sampling volume of blood specimens and quicker analysis.…”