| Infectious bovine rhinotracheitis?IBR?is a contagious infectious disease caused by Infectious bovine rhinotracheitis virus?IBRV?.The disease is mainly caused fever,severe upper respiratory symptoms by cattle,cow abortion,stillbirth,and Inflammation of the reproductive system and calf encephalitis.In China,the prevalence of IBRV has been increasing,which has caused heavy economic losses to China's cattle rearing.How to effectively prevent and control Infectious bovine rhinotracheitis has become a iscritical issue of the cattle rearing.Because the diagnosis of Infectious bovine rhinotracheitis is difficult,the conventional detection technology is complex,be time-consuming,be strenuous and its result is not accurate.The colloidal gold immunochromatography technique is simple,rapid,sensitive,and stable.Intuitive and other advantages,it can provide a new way for the detection of Infectious bovine rhinotracheitis.In this study,DNAstar software was used to analyze,predict,and screen out gD antigens with high antigenic index and good hydrophilicity?gDa,gDb,gDc?for Infectious Bovine Rhinotracheitis.Gene sequence was synthesized to obtain the fusion gene pET-28a?+?-gDabc,which was transformed into E.coli competent cell BL21?DE3?plysS.The expression was induced by adding 0.5mmol/L IPTG and the expressed product was detected by SDS-PAGE.The molecular weight of protein is 37 kDa,it is consistent with the theory was calculated.Weartn-blot analysis showed that pET-28a?+?-gDabcbc was highly expressed recombinantly in E.coli.The purified fusion protein gDabcbc was used as the coating antigen for the test line,the Staphylococca A protein antibody as the control line was streaked on the nitrocellulose membrane,and the colloidal gold-SPA was sprayed onto the Glass Fibre Mats.Established an immuneochro-matographic method for the antibody colloidal gold in bovine infectious rhinotracheitis;IBRV standard positive serum was detected by the established colloidal gold test paper,Red reaction line appeared on the check line and quality control line.When the negative serum was detected,only the quality control line appears red stripe.This indicates that the test strip showed a positive reaction when the positive serum was detected,and the negative serum was detected when the negative serum was detected,the results are effective;test strips can detect the most appropriate antigen concentration of Bovine Infectious Rhinotracheitis antibody is 0.6 mg/mL;test strips to detect bovine infectious rhinotracheitis negative serum,Bovine Paratuberculosis positive serum,Bovine viral diarrhea positive serum,bovine brucellosis positive serum,and positive serum of foot-and-mouth disease in cattle were negative;IBRV was detected using a test strip stored at room temperature for 6 months and a test strip stored at 4°C for 6 months.The test results of negative serum and positive serum were consistent with those before storage.The prepared test strip has the advantages of sensitivity and speciality,good stability,the results within 15 min,and the operation is simple and can be used for clinical diagnosis. |
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