Diagnosis of Pasteurella Multocida and Mannheimia Haemolytica Infections in Cattle Using Lateral Flow Immunochromatographic Assay

Abstract

This study was mainly designed to advance and recover the control measures of Bovine respiratory disease (BRD) via accelerating the field identification of some BRD causing pathogens. A simple rapid field test was developed mainly to identify Pasteurella multocida and Mannheimia haemolytica, both are highly associated with BRD. A full identified Pasteurella multocida and Mannheimia haemolytica cultures were earned from Microbiology Department, Faculty of Veterinary Medicine, Cairo University, Egypt. Species-specific bacterial antigen of outer membrane protein of each bacterial strain was prepared separately for M. haemolytica and P. multocida. Specific polyclonal antibodies were prepared by amplifying each one in guinea pigs and rabbits. Pathogens-specific lgG were purified using caprylic acid technique. Pathogens-specific lgG titers were measured by ELISA. The colloidal gold nanoparticles were conjugated to the rabbit mono specific polyclonal lgG antibody and did not conjugate to guinea pig specific polyclonal lgG. The particles were then utilized to develop the Lateral flow kits (LFK) which is a rapid assay for detection for P. multocida and M. haemolytica in clinical samples when Compared to the conventional Bacteriological tests. Results: Percentage of accuracy, specificity and sensitivity of LFK kits recorded 93, 94.4 and 90.4%, respectively, for P. multocida and 91.5, 94.3and 84.7%, respectively, for Mannheimia haemolytica.