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

• Published in: Journal of virological methods Vol. 222; pp. 164 - 169
• Main Authors: 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
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 15.09.2015
• Subjects: Animals; Antibodies, Viral - blood; Antibody detection; Herpesvirus 1, Suid - immunology; ICS; Immunochromatography - methods; PRV; Pseudorabies - diagnosis; Pseudorabies virus; Sensitivity and Specificity; Swine; Temperature; Time Factors; Viral Envelope Proteins - immunology; ICS; Antibody detection; PRV
• ISSN: 0166-0934; 1879-0984
• DOI: 10.1016/j.jviromet.2015.06.016

•An ICS was developed with high specificity, reproducibility and stability.•This method can be used for detecting PRV wild-type viruses.•The challenge of PRV in swine used to evaluate the result of the ICS. 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).