Development of a Colloidal Gold Immunochromatographic Strip Assay for Rapid Detection of Bovine Rotavirus.

Publisher: Mary Ann Liebert, Inc Country of Publication: United States NLM ID: 8801552 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1557-8976 (Electronic) Linking ISSN: 08828245 NLM ISO Abbreviation: Viral Immunol Subsets: MEDLINE

Original Publication: New York, NY : Mary Ann Liebert, Inc., c1987-

Antibodies, Viral/*chemistry ; Cattle/*virology ; Diarrhea/*diagnosis ; Gold Colloid/*chemistry ; Rotavirus/*isolation & purification ; Rotavirus Infections/*diagnosis; Animals ; Antibodies, Immobilized/chemistry ; Antibodies, Monoclonal/chemistry ; Antibodies, Monoclonal/immunology ; Antibodies, Monoclonal/isolation & purification ; Antibodies, Viral/immunology ; Antibodies, Viral/isolation & purification ; Antigens, Viral/immunology ; Capsid Proteins/immunology ; Diarrhea/veterinary ; Diarrhea/virology ; False Negative Reactions ; False Positive Reactions ; Immunoassay/instrumentation ; Rotavirus/immunology ; Rotavirus Infections/veterinary ; Rotavirus Infections/virology ; Sensitivity and Specificity ; Time Factors

Bovine rotavirus (BRV) is one of main pathogens responsible for diarrhea, fever, and vomiting. In this study, we developed a colloidal gold immunochromatographic test strip for detecting BRV according to the principle of double-antibody sandwich. The monoclonal antibodies (mAbs) and polyclonal antibodies (pAbs) were prepared and purified. On the strip, the purified mAbs labeled with the colloidal gold were used as the detector, and the goat anti-mouse antibodies and purified pAbs were coated on the nitrocellulose membranes as the control line and the test line, respectively. We optimized different reaction conditions, including the amount of mAbs, the pH of colloidal gold solution, coating solution, blocking solution, sample pad treatment solution, antibody concentration in control line, and antibody concentration in detection line. In specificity assay, the strip had high specificity in detecting BRV. No cross-reaction was observed in detecting other viruses. The detection sensitivity of the strip was found to be 1 × 10 ³ TCID 50 /0.1 mL. Two hundred twenty clinical samples were detected with the strip compared to reverse transcription-polymerase chain reaction. No false-negative or false-positive results were found, and the results obtained by the two methods were similar. In conclusion, we developed a novel immunochromatographic strip to rapidly detect BRV. The strip developed exhibited high sensitivity and specificity for BRV detection. It could be a rapid, convenient, and effective method for the rapid diagnosis of BRV infection in the fields.

Keywords: bovine rotavirus; colloidal gold immunochromatographic strip; monoclonal antibodies

0 (Antibodies, Immobilized)
0 (Antibodies, Monoclonal)
0 (Antibodies, Viral)
0 (Antigens, Viral)
0 (Capsid Proteins)
0 (Gold Colloid)
0 (VP6 protein, Rotavirus)

Date Created: 20191010 Date Completed: 20200727 Latest Revision: 20200727

20240104

10.1089/vim.2019.0071

31596683