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PROFIL IMUN RESPON TERHADAP RABIES DAN ANALISIS GENETIKA GEN PENYANDI GLIKOPROTEIN VIRUS RABIES ISOLAT BALI


Oleh : NI LUH DARTINI | -
Bidang Ilmu : Bioteknologi | Tahun Penelitian : 2011

Download PROFIL IMUN RESPON TERHADAP RABIES DAN
Download PROFIL IMUN RESPON TERHADAP RABIES DAN
Download PROFIL IMUN RESPON TERHADAP RABIES DAN


ABSTRAK

Ni Luh Dartini, NIM, 0990861006. “Profil Imun Respon Terhadap Rabies dan Analisis Genetika Gen Penyandi Glikoprotein Virus Rabies Isolat Bali”. Di bawah bimbingan IGNK Mahardika dan AA Gde Putra. Penelitian ini dilakukan untuk mengetahui profil imun respon anjing yang di vaksinasi dengan vaksin Rabivet Supra92 dan vaksin Rabisin, dengan satu kali dosis vaksinasi, dalam kondisi lapangan. Tujuan kedua adalah uji banding kit ELISA Pusvetma dan PlateliaII Bio-rad, dan terakhir adalah analisis terhadap gen penyandi glikoprotein virus rabies isolat Bali. Sebanyak 53 sampel serum anjing yang belum divaksinasi, 463 sampel serum anjing pasca vaksinasi dengan vaksin Rabivet supra92 dan 267 sampel serum anjing pasca vaksinasi dengan vaksin Rabisin, di Provinsi Bali, diambil sejak Desember 2008 sampai Maret 2011. Lima isolat virus rabies dari otak anjing di amplifikasi dengan RT-PCR dan dilakukan sekuensing di Eijkman Institut. Isolat virus rabies tersebut tiga berasal dari Bali, satu dari Flores, dan satu dari Sulawesi. Sampel otak anjing itu dikoleksi Tahun 2009. Semua sampel serum dari lapangan diuji dengan ELISA kit produksi Pusvetma Surabaya. Data hasil uji banding kit ELISA dianalisis berdasarkan agreement between test (Kappa), dan data hasil uji gen penyandi glikoprotein dianalisis menggunakan program MEGA4. Hasil penelitian menunjukkan bahwa persentase anjing yang mempunyai titer antibodi protektif yang divaksinasi dengan vaksin Rabivet Supra92 dan Rabisin tidak berbeda nyata (P=0,971). Namun demikian, titer antibodi terhadap rabies pada anjing yang divaksinasi dengan vaksin Rabisin sangat nyata lebih tinggi dibandingkan dengan yang divaksin Rabivet Supra92 (P=0,000). Dalam penggunaan kedua vaksin tersebut, persentase antibodi protektif tertinggi diperoleh pada periode lebih dari tiga bulan sampai dengan enam bulan pasca vaksinasi. Selanjutnya, kit ELISA Pusvetma mempunyai kekuatan kesepakatan yang baik dengan Kit Baku PlateliaII dengan nilai kappa sebesar 0,68 (Substantial agreement). Sedangkan analisis fragmen gen penyandi glikoprotein virus rabies isolat Bali menunjukkan bahwa virus rabies isolat Bali berada dalam satu kluster dan mempunyai jarak genetik paling dekat dengan virus rabies isolat Flores.


Kata Kunci: Respon imun, Rabies, vaksinasi, rabivet supra92, rabisin, ELISA, gen penyandi glikoprotein.


ABSTRACT

Ni Luh Dartini, NIM, 0990861006. “Profile Immune Response to Rabies and Analysis of Glycoprotein Gene Rabies Virus Isolate Bali”. Under supervision of IGNK Mahardika dan AA Gde Putra. This study was conducted to determine the profile of immune responses in vaccinated dogs with Rabivet Supra92 and Rabisin vaccines. The second objective was to compare the ELISA test kit produced by Pusvetma Surabaya with a standard validated kit PlateliaII (Bio-rad). The last was to analyze the fragment gene encoding the glycoprotein of rabies virus. The number of samples was 53 dog sera that have not been vaccinated, 463 dog sera after vaccination with Rabivet supra92, and 267 dog sera after vaccination with the vaccine Rabisin. The sera were collected in Bali Province from December 2008 until March 2011. Five rabies virus from dog brain have been amplified using RT-PCR and sequenced in Eijkman Institute. The origin of dog samples was 3 from Bali, one from Flores, and one from Sulawesi. All brain samples were collected in 2009. All serum samples from the field were tested by ELISA kit Pusvetma. The agreement of the kits (Pusvetma and Platelia II rabies kits Bio-rad) was analyzed based on Kappa value, and the sequence data of the gene encoding glycoprotein were analyzed using MEGA4 program. The results showed that the percentage of dogs that have a protective antibody that were vaccinated with Rabisin and Rabivet Supra92 vaccines, was not significantly different (P = 0.971). However, antibody titers against rabies in dogs vaccinated with Rabisin vaccine were significantly higher than those that were vaccinated with Rabivet Supra92 (P=0.000). Both vaccines generated the highest percentage of protective antibodies in the period of more than three to six months post-vaccination. Furthermore, ELISA kit Pusvetma has a substantial agreement with the standard Platelia kit with a kappa value of 0.68. While the analysis of gene fragment that codes for the glycoprotein of rabies virus showed that the Bali’s rabies viruses were closely related and formed a single cluster phylogenetically, and have the closest genetic distance with the rabies virus from Flores.


Keywords: Rabies, immune response, dog, vaccination, Rabivet Supra92, Rabisin, ELISA, glycoprotein gene.