Detection of Immunoglobulin Geneheavy Chain Binding Protein in Eimeria tenella Collected from Yogyakarta Using One Step Reverse Transcriptase PCR
Keywords:IHC-BiP, Eimeria tenella, reverse transcriptase PCR, Yogyakarta
Coccidiosis is an infection disease caused by Eimeria tenella in chicken intestine. The disease causes the economic lost in poultry industry. This disease is very difficult to be controlled for some reasons, e.g.the lack of accurate diagnostic tools, expensive vaccinationcost, and coccidiost at resistance in Eimeria. Immunoglobulin heavy chain binding protein (IHC-BiP) is an immunogenic agent for internal parasite which involves 70 families of Eimeria in the heat shock protein. Gene sequence of the binding protein(IHC-BiP) was found in E. tenella and E. maxima from United Kingdom, however there is no information about this gene in Eimeria from Indonesia. This research was aimed to find IHC- BiP gene in four developmental stages of E. tenella collected from Yogyakarta. Samples were collected from the field and propagated in 50 chickens. Each chicken was infected with 10.000 E. tenella oocysts. The oocyst of E. tenella from propagation were then sporulated and prepared for sporocyst and sporozoite stages. Total RNAs were isolated from each stage of E. tenella development (unsporulated oocyst, sporulated oocyst, sporocyst, and sporozoite) and the gene was detected by one step reverse transcriptase PCR(RT-PCR). The result showed that the IHC-BiP gene was only found in sporocyst stage of E. tenella and did not in other stages. The gene has size of 714 bp. This study suggests that the IHC-BiP protein of E. tenella from Yogyakarta is produced during sporulation, especially in sporocyst stage.