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Development and clinical evaluation of a new gold-immunochromatographic assay for the detection of antibodies against field strains of pseudorabies virus

Guo, Dian-lei, Pan, Qi-wei, Li, Kun-peng, Li, Jun-qing, Shen, Han-wei, Wang, Xiang-ling, Zhang, Xun-yun, Li, Xue-song, Fu, Fang, Feng, Li, Li, Xi
Journal of virological methods 2015 v.222 pp. 164-169
gold, Suid alphaherpesvirus 1, immunoglobulin G, blood serum, immunoaffinity chromatography, antibodies, storage temperature, analytical kits, staphylococcal protein A, clinical examination, glycoproteins, swine
An immunochromatographic strip (ICS) was developed for the detection of swine antibodies against glycoprotein E (gE) in Pseudorabies Virus (PRV). In this test, Staphylococcal Protein A (SPA) labeled with colloidal gold was dispensed on a conjugate pad as the detector. Purified PRV-gE and pig-IgG were blotted on a nitrocellulose membrane for the test (T) and control lines (C), respectively. If the tested serum contains IgG antibodies against PRV-gE, the IgG will interact with the colloidal gold-SPA to form a complex (gold-SPA-swine IgG). The complex will react with the immobilized PRV-gE on the T line and the Pig-IgG in the C line of the ICS to form two visible red bands. If there is no IgG antibody against PRV-gE in the sample serum, only the C line will be visible. The ICS was capable of specifically detecting PRV-gE antibody within 5min, and its stability and reproducibility were quite good after storage at 4°C and use within 4 months. Using an IDEXX Pseudorabies Virus gE Antibody Test Kit (IDEXX PRV gE Ab test) as a reference, the relative specificity and sensitivity of the ICS were determined to be 81.6% and 90.7%, respectively. Furthermore, there was a good agreement between the results obtained by the commercial product and the ICS (kappa=0.7289).