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

Abstract

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 × 103 TCID50/0.1 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.