%T Development of Rapid Detection Kit for Necrotic Enteritis Disease in Poultry using Protein A Agglutination
%N 1
%X Necrotic enteritis causes significant losses in the global poultry industry, necessitating accurate diagnosis for effective intervention. This study aimed to develop a diagnostic tool for detecting necrotic enteritis in poultry based on the presence of Clostridium perfringens (C. perfringens) Alpha-toxin in poultry feces. The reagent of the detection kit was developed by conjugation of IgG against C. perfringens toxin and Staphylococcus cells containing protein A. The IgG antibody was derived from an 8-month-old thin-tailed male sheep immunized with purified 2 ml of C. perfringens Alpha-toxin. Sensitivity assays were carried out to determine the detection limit, while Escherichia coli (E. coli) and Salmonella enteritidis (S. enteritidis) were used to identify specificity. A purified Alpha-toxin with a protein concentration of 2.8 mg/ml and a specific molecular weight of 43 kDa was successfully obtained. A strong reaction of the hyperimmune antibody (IgG) was also detected in the thin-tailed male sheep serum. The developed rapid detection kit in this study indicated C. perfringens Alpha-toxin with a lower concentration (12 ng/ml). Agglutination reactions could differentiate positive control from negative without significant cross-reactivity towards other bacteria (S. enteritidis and E. coli). © The Author(s) 2024
%O Research and Diagnostic Unit, PT. Medika Satwa Laboratoris, Bogor, 16166, Indonesia; Doctoral Program in Biomedical Science, Faculty of Medicine, Universitas Indonesia, Jakarta, 10430, Indonesia; School of Veterinary Medicine and Biomedical Sciences, IPB University, Bogor, 16680, Indonesia; Department of Medical Pharmacy, Faculty of Medicine, Universitas Indonesia, Jakarta, 10430, Indonesia; Department of Animal Diseases Sciences and Veterinary Public Health, Faculty of Veterinary Medicine, University of Nusa Cendana, Kupang, 85001, Indonesia; Department of Clinical Microbiology, Faculty of Medicine, Universitas Indonesia, Jakarta, 10430, Indonesia; Department of Animal Disease and Veterinary Health, School of Veterinary Medicine and Biomedical Sciences, IPB University, Bogor, 16680, Indonesia
%J World's Veterinary Journal
%I Scienceline Publication, Ltd
%L eprints1213
%P 8-14
%D 2024
%R 10.54203/scil.2024.wvj2
%V 14
%A R.S. Kurnia
%A C.M.H. Nugroho
%A O.S.M. Silaen
%A M.A. Putra
%A V.A. Rahmi
%A A. Amaliah
%A R.W. Hakim
%A M.U.E. Sanam
%A A. Soebandrio
%A . Safika
%A A. Indrawati
%K Clostridium toxin; immunoglobulin G; immunoglobulin G antibody; protein A, agglutination test; animal experiment; antibody production; Article; bacterial cell; bacterium detection; Clostridium perfringens; concentration (parameter); conjugation; controlled study; cross reaction; Escherichia coli; feces analysis; female; immunization; intermethod comparison; limit of detection; molecular weight; necrotizing enteritis; nonhuman; polymerase chain reaction; poultry; protein purification; Salmonella enterica serovar Enteritidis; sensitivity analysis; sensitivity and specificity; sheep; Staphylococcus