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All Journal JURNAL BIOLOGI INDONESIA Jurnal Kedokteran Hewan
Yuniarto, Ichwan
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Biochemistry, Genetics & Molecular Biology Immunology & microbiology Medicine & Pharmacology Veterinary

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IDENTIFIKASI PROTEIN DAN SELEKSI ISOLAT TRYPANOSOMA EVANSI BERSIFAT IMUNOGENIK UNTUK KANDIDAT PENGEMBANGAN IMUNOASAI Subekti, Didik T.; Yuniarto, Ichwan
JURNAL BIOLOGI INDONESIA Vol 14, No 1 (2018): JURNAL BIOLOGI INDONESIA
Publisher : Perhimpunan Biologi Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/jbi.v14i1.3669

Abstract

ABSTRACTSurra is a parasitic disease caused by T. evansi and causing high economic losses in Indonesia. Some isolates have been isolated from several areas experiencing outbreaks of Surra in Indonesia. The isolates have been reported to have a diversity of protein profiles based on SDS PAGE (sodium dodecyl sulphate polyacrilamide gel electrophoresis). The research aims to identify immunogenic protein from each isolate and make the selection of T. evansi isolates which is potential as a source of antigens for immunoassay development. Each isolates were obtained to be purified from blood and then the protein was isolated. The proteins were run onto polyacrylamide gel electrophoresis and visualized by Coomassie blue. Another electrophoresis results were transferred onto a nitrocellulose membrane for immunoblotting. The results showed that the immunogenic proteins that consistently detected among nine T. evansi isolates of Indonesia are 100, 90, 85, 76-80, 70, 65, 55, 49-52, 44-46, 40, 34-36, 31 -33 kDa. Among the nine T. evansi isolates, N372 isolate was selected as a candidate for immunoasay development, especially ELISA. Immunogenic proteins were specifically found on the N372 isolate are 85, 70, 65, 49-52, 44-46, 34-36, 31-33, 24-28, 15-20 kDa.Keywords : Trypanosoma evansi, immunoblotting, protein profiles, immunogenic protein
PERBANDINGAN PENGGUNAAN KONJUGAT ANTIBOVINE IGG-HRP DAN PROTEIN A/G-HRP DENGAN BEBERAPA LARUTAN PENGENCER SERUM PADA ELISA UNTUK DETEKSI SURRA PADA SAPI DAN KERBAU Subekti, Didik T.; Yuniarto, Ichwan
JURNAL BIOLOGI INDONESIA Vol 13, No 1 (2017): JURNAL BIOLOGI INDONESIA
Publisher : Perhimpunan Biologi Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/jbi.v13i1.3093

Abstract

ABSTRACTSurra was reported in various types of animals both livestock, pets and wildlife. ELISA (enzyme linked immunosorbentassay) is a sensitive and specific diagnosis technique for Surra detection. The use of speciesspecific conjugate (anti bovine IgG-HRP) has limited while protein A/G can be used for a wide variety of animalspecies. This study aimed to evaluate the initial application of the protein A/G-HRP compared with antibovine IgG-HRP using standard samples using four (4) types of diluent buffer. The standard serum samples (23serum) consist of positive and negative sera from bovine (cattle and buffalo) were reacted with Surra antigenon microplate. Positive and negative serum was diluted with different diluent buffer, namely PBS-Tween20(PBST), RBA (RedBuff A), RBB (RedBuff B) and LC (Low Cross). Results of ELISA using protein A/G-HRPshowed absorbance values reduced 36.16% - 69.30% compared to the anti-bovine IgG-HRP. The percentagereduction of PBST, RBA, RBB and LC, were 51.76%; 56.64%; 36.16% and 69.30% respectively. The use ofprotein A/G-HRP and fourth diluent buffer can reduce antigen - antibody bonding with a weak affinity whichlowers the absorbance value of ELISA Surra.Keywords : Protein A, Protein G, Surra, ELISA, Trypanosoma evansi
COMPARISON OF POLYPEPTIDE PROFILE OF Trypanosoma evansi ISOLATES FROM INDONESIA AND THEIR RELATION TO BIOTYPE AND SENSITIVITY TO TRYPANOCIDAL Yuniarto, Ichwan; Subekti, Didik T; Cahyaningsih, Umi; Satrija, Fadjar
Jurnal Kedokteran Hewan Vol 12, No 2 (2018): June
Publisher : Universitas Syiah Kuala

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21157/j.ked.hewan.v12i2.11486

Abstract

This study aimed to determine whether the variant or biotype of Trypanosoma evansi can be seen from their polypeptide profiles using 12%sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS PAGE) stained with Brilliant Blue Commasie. The results generally showed thatthe molecular weight (MW) of polypeptides from nine isolates from East Java, Central Java, Banten, South Kalimantan, Central Kalimantan, andLampung provinces were in the range of 85.46 to 15.76 kD and each isolate has different polypeptide profile. Isolates A13 and A14 were isolatedfrom the same place but have different polypeptide profiles. Likewise, isolates S13 and S18 also have different polypeptide profiles despite beingisolated from the same place at the same time. On the other hand, isolate 372, 87, and 06 have different protein profiles but was classified in thesame biotype namely biotype I. Generally, the difference in protein profile actually more related to the biological diversity of the metabolism ofeach Trypanosoma evansi isolate from Indonesia.
Co-Authors Didik T Subekti Didik T. Subekti Umi Cahyaningsih