A novel immunochromatographic strip for rapid and field-deployable detection of pigeon adenovirus type 1

Pigeon adenovirus type 1 (PiAdV-1) is a causative pathogen of juvenile pigeon diarrhea disease, leading to huge economic losses to the global pigeon industry. Rapid detection on the poultry farm is helpful for the disease control. In this study, the Hexon and Fiber-2 proteins of PiAdV-1 were recombinantly expressed in a prokaryotic expression system using vector pET-28a and used for the immunization of the BALB/c mice. Two monoclonal antibodies (mAbs), 1H2 and 2F1, targeting the Hexon and Fiber-2 proteins respectively were successfully screened by the traditional method of hybridoma fusion. An immunochromatography test strip (ICS) was developed by using colloidal gold-labeled 1H2 and used for the rapid detection of PiAdV-1. The characteristic analysis revealed that the ICS displayed excellent sensitivity and specificity with a detection limit of 5.76 × 10³ copies/μL and showed no cross-reactivity with common pigeon viruses including PiAdV-2, pigeon paramyxovirus, pigeon circovirus, and pigeon rotavirus. In addition, the ICS was validated by comparison with a polymerase chain reaction (PCR) for the 300 samples detection. The diagnostic positive rates of ICS and PCR were 22.67% (68/300) and 25.33 (76/300) respectively. The agreement rate was 97.34% between results obtained with ICS and PCR on 300 field fecal samples. Therefore, the ICS will be helpful for the rapid and effective detection of PiAdV-1 in clinical fecal samples.