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Abstract
BackgroundInfectious coryza (IC) is an acute respiratory disease of growing chickens and layers caused by Avibacterium paragallinarum. The development of tools that allow rapid pathogen detection is necessary in order to avoid disease dissemination and economic losses in poultry. An Av. paragallinarum-specific Ma-4 epitope of the TonB-dependent transporter (TBDT) was selected using bioinformatic tools in order to immunize a BalbC mouse and to produce monoclonal antibodies to be used in a lateral flow test (LFT) developed for Av. paragallinarum detection in chicken nasal mucus samples.ResultsThe 1G7G8 monoclonal antibody was able to detect TBDT in Av. paragallinarum cultures (serogroups: A, B and C) by Western blot and indirect ELISA assay. Consequently, we developed a self-pairing prototype LFT. The limit of detection of the prototype LFT using Av. paragallinarum cultures was 1 × 104 colony-forming units (CFU)/mL. Thirty-five nasal mucus samples from chickens suspected of having infectious coryza were evaluated for the LFT detection capacity and compared with bacterial isolation (B.I) and polymerase chain reaction (PCR). Comparative indicators such as sensitivity (Se), specificity (Sp), positive predictive value (PPV), negative predictive values (NPV) and the kappa index (K) were obtained. The values were 100.0% Se, 50% Sp, 65.4% PPV, 100% NPV, and 0.49 K and 83.9% Se, 100% Sp, 100% PPV, 44.4% NPV, and 0.54 K for the comparison of the LFT with B.I and PCR, respectively. Additionally, the LFT allowed the detection of Av. paragallinarum from coinfection cases of Av. paragallinarum with Gallibacterium anatis.ConclusionsThe results indicate that the self-pairing prototype LFT is suitable for the detection of TBDT in Av. paragallinarum cultures as well as in field samples such as nasal mucus from Av. paragallinarum-infected chickens. Therefore, this prototype LFT could be considered a rapid and promising tool to be used in farm conditions for Av. paragallinarum diagnosis.
Highlights
Infectious coryza (IC) is an acute respiratory disease of growing chickens and layers caused by Avibacterium paragallinarum
The most common clinical signs are serous or mucous nasal exudates, sneezing, swelling of infraorbital sinuses, facial edema and conjunctivitis [2]. All these clinical signs caused by Av. paragallinarum have been associated with economic losses in the poultry industry [2] that highlight the necessity of developing reliable tools for Av. paragallinarum detection
Monoclonal antibody characterization Two hybridoma clones, 1G7G8 and 3A3D8, were selected based on their reactivity against the peptide (Ma-4); 1G7G8 and 3A3D8 produced monoclonal antibodies, which were characterized as IgG1 (k) and IgG2b (k) isotypes, respectively
Summary
Infectious coryza (IC) is an acute respiratory disease of growing chickens and layers caused by Avibacterium paragallinarum. The most common clinical signs are serous or mucous nasal exudates, sneezing, swelling of infraorbital sinuses, facial edema and conjunctivitis [2] All these clinical signs caused by Av. paragallinarum have been associated with economic losses in the poultry industry [2] that highlight the necessity of developing reliable tools for Av. paragallinarum detection. The use of a lateral flow test for detection of Av. paragallinarum is an alternative method that is not demanding and is an performable task. This method requires the identification of a specific epitope in the target protein that is detected through the use of a monoclonal antibody
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