Development of a Lateral Flow Assay for Rapid Detection of Bovine Antibody toAnaplasma marginale

Abstract

A rapid lateral flow assay for detection of bovine antibody to Anaplasma marginale was developed. The assay used a recombinant peptide of major surface protein 5 as the antigen and a monoclonal antibody specific for bovine IgG(1) conjugated with colloidal gold beads for detection. Serum and anticoagulated blood samples were obtained from cattle in an area where anaplasmosis was endemic. The samples were selected based on positive identification of the organism in blood smears. The unclotted blood samples were used for PCR determination of the presence of A. marginale while the sera were tested by a commercial competitive enzyme immunoassay (CELISA) and by the lateral flow assay (LFA). Similar samples, collected at a Canadian sales barn, were tested by the CELISA and LFA and 10% were tested by PCR for the presence of A. marginale nucleic acid. In addition, stored serum samples from a second endemic area were tested by CELISA and LFA. Of the 114 smear positive samples, all were positive by CELISA and LFA. All samples were also positive by PCR. Samples from Canadian sources (n=524) were negative in the CELISA but 11 sera gave false positive reactions in the LFA. All samples tested were PCR negative. Of 113 samples from herds with anaplasmosis, 53 were positive in the CELISA and 50 were LFA positive.

A Review of Bovine Anaplasmosis

“…Canadian researchers from Canadian Food Inspection Agency have recently developed a lateral flow assay (LFA) for the rapid detection of bovine antibody to A. marginale (Nielsen et al., 2008). The assay uses a recombinant peptide of MSP5 as the antigen and a monoclonal antibody specific for bovine IgG 1 conjugated with colloidal gold beads for detection.…”

Lateral flow (immuno)assay: its strengths, weaknesses, opportunities and threats. A literature survey

“…Lateral flow (immuno)assay is a technology that is currently widely applied [5][6][7][8][9][10][11][12][13][14][15][16]; however, to our knowledge no in-depth survey of recent literature is available. Here we present a survey of development reports presented in the scientific literature, mainly during the last decade, where a selection of those applications is mentioned that are real one-step dry reagent assays with coloured or luminescent response.…”

“…The first labels were the enzymes used for enzyme immunoassays [4], but they were replaced soon by particulate labels [1]. Nowadays when a new method is presented for LFIAs, the performance of the new system is often referenced to the performance of enzyme immunoassays [5][6][7][8].…”

Investigating Membrane Morphology and Quantity of Immobilized Protein for the Development of Lateral Flow Immunoassay

“…In recent years, lateral flow diagnostic kits have been widely introduced in the biomedical and healthcare fields [1][2][3] due to their high sensitivity, rapid testing, low manufacturing cost, and simple operating procedures. [4] A large number of diagnostic kits are available for the detection of certain infectious diseases, [5][6][7] verification of food hygiene, [8,9] and testing of fertility.…”