Development of Enzyme-Linkage Immunosorbent Assay Against Type B of Clostridium Botulinum: A Preliminary Study

Abstract

ABSTRAKNeurotoksin Clostridium botulinum (BoNTs) merupakan salah satu penyebab kerugian ekonomi bagi industri peternakan. Kerugian ini dapat sebagai akibat langsung ketika ternak keracunan BoNTs atau secara tidak langsung ketika produk asal ternak terkontaminasi BoNTs sehingga produk tersebut dilarang untuk dipasarkan. Untuk itu perlu dilakukan pengontrolan rutin penyebaran BoNTs di lingkungan peternakan dan industri hasil ternak. Salah satu caranya adalah dengan melakukan deteksi rutin terhadap keberadaan BoNTs. Salah satu metode deteksi yang relatif cepat dan akurat adalah metode enzyme linkage immunosorbant assay (ELISA). Pada penelitian ini dilaporkan hasil pengembangan ELISA, menggunakan antibodi poliklonal produk lokal terhadap BoNTs-B. Antibodi dihasilkan dari enam mencit Balb/c dengan metode imunologi standar. Mencit diimunisasi 3 kali selama 8 minggu dengan toksoid C. Botulinum tipe B komersial dengan dosis 100 ng per ekor per injeksi. Antibodi yang dihasilkan dimurnikan dengan kombinasi metode presipitasi ammonium sulfat 50% (w/v) dan kolom protein A. Hasil studi pendahuluan ini menunjukkan bahwa metode ELISA yang dikembangkan mampu mendeteksi toksin Clostridium botulinum tipe B hingga 1,0 ng/ml. Kata kunci: toksin C. botulinum, ELISA, antibodi poliklonal ABSTRACTClostridium botulinum neurotoxin (BoNTs) is one of the causes of economic loss in the livestock industry. This economic loss would be as a direct result when animals poisoned by BoNTs or indirectly when the livestock products are contaminated by BoNTs, which end up with the products are banned by authority. Therefore a routine surveillance of BoNTs in the farm and in livestock product processing industry is urgently needed. One of the most relatively quick and accurate methods to perform a routine detection of the presence of BoNTs is enzyme-linkage immunosorbant assay (ELISA). In this article we describe the results of the development of ELISA, using polyclonal antibodies against BoNTs-B produced locally. Antibodies were generated from six Balb/c mice with standard immunological methods. Mice were immunized three times for a period of 8 weeks with a commercial type B Clostridium botulinum toxoid at a dose of 100 ng per mouse per injection. The resulting antibody was purified by a combination of ammonium sulfate precipitation 50% (w/v) technique and a protein A column method. The results of this preliminary study indicated that the developed ELISA method capable of detecting type B Clostridium botulinum toxin up to 1.0 ng/ml.

Utilization of Polyclonal Antibodies Produced in Local Horses (Equus Caballus) as a Resource for Development of Elisa Conjugate to Detect Hepatitis B Virus (Hbv) Surface Antigens

“…Quantitative data regarding the ELISA tests were tabulated and analyzed according to Drouet et al (2003), and Depamede and Kisworo (2011).…”