Pola migrasi RNA genome rotavirus group A dengan g-genotype

DWIJA, Ida Bagus Nyoman Putra, dr. Abu Tholib Aman, M.Sc., Ph.D., Sp.MK

2009 | Tesis | S2 Bioteknologi

Rotavirus merupakan penychab diare non hakterial paling dominan yang ditemuì dìmasyarakat dan menyebabkan beban kesehatan yang besar baik di negara maju maupun negara berkembang. Rotavinis termasuk kedalam family Reoviridae, genorrr berupa dsRNA yang cerdiri atas 11 segn’tn yang mengkode 6 protein structural dan 6 protein non-strukural. Bedasarkan ‘P6 rntavirus dikelompokkan dalam 7 group A-G, group A rriernpakan penyebab utama diare pada bayi dan anak-anak urnur < 5 tahun. Dua kapsid bagian luar VP4 (P-type) dan VP? (C-type) mampu menginduksi munculnya respon imun. 61-4. (39 dan P[4J, P[6], PiS] mewpakaìð strain dominan yang ditemukan dimasyarakat. Pola migrasi genorne rotavinis pacía gel elektroforesis memiliki pola 4-2-3-2, berdasarkan migrasi segmen 10 dan 11 dikelompokkan menjadi pola panjang dan pendek Tingginya variasi strain dan pela elekimforesis memerlukan suatu studi untuk mengetahui hubungan strain dan pola elektroforesis yang dítemukan. Penelitian ini menggunakan sainpel feses thui RSU Kola Yogyakarta dan daerah lain di Indonesia. Ro(avirus group A ditenlukan dengan ELISA (WEJAIM Rotavinis, Daim Cyromation), ekstraksi RNA dilakukan dengan rnenggunakan Thzol LS (Tnvitrogen). Genotiping dengan RT-PCR menggunakan primer spesifik untuk 6- type dan elektroforesis sesuai slaudar WHO. Lul.ektroferotipe cliaruali dengan poliakrilainid gel elektroforesis (PAGE) dan Roche dan pewarnaan menggunakan perak nitrat Selarna 6 Inflan penelitian terkumpul 85 sanipel feries dan bayi ci tahun, dan RSU Kola Yogyakarta. ELISA dilakukan terhIap 84 s.ampel, 44(52%) positif rocavirus. (Jenotyping dapat dilalcukan pacía 43 sampel, 01 ditemukan parla 34 (79%) sampel $ 02 path 9 (21%). PAGE terhadap 43 sampel rnenunjukkan Gi memiliki pola panjang, sedang 62 pola pendek. Jika dibandìngkan dengan strain daii daerah lain ditemukan 4 pola berbeda, Dapat disimpulkan strain (31 dan (39 memilild pela panjang, sethmg 62 dan G3 memiliki pola pendek

Rotavinis is the most dominant cause of non bacterial diarrhea encountered in the community aoci causing major hea]th burden both in developed and developing country. Rotavirus belong to the family of Reo viridue, a dsRNA genome consisting of 1 segmenLs thai. encode 6 structural proteins and 6 non- structural proteins. Based on VP6 rolavirus grouped in a 7 group A- G, group A is the main cause of diarrhea in infants and young children aged <5 years. Two outer capsid VP4 (P-type) and VP’) (0-type) can induce the emergence of an immune Tesponse. 01-4, G9 and P [4], p [6J, P igi is the most common strain found in community. Migration pattern of rotavirus group A genome in the gel electrophoresis has a 4-2-3-2 pattern, based on the migration of segments 10 and I 1,rotavirus are grouped into short and long patterns. The high sirain variation and patterns of electrophoresis requires a smdy to determine correlation between strain and electrophoresis patterns of rotavinis strains. This study used fecal samples from RSU Kota Yogyakarta and various regions in Indonesia. Rotavirus group A is determined by ELISA (Rotavirus IDEIATh, Daco Cytomation), extraction is done by using Trizol LS (Invitrogen) method. Genotyping by RT-PCR uses specific primers for VP7 gen, according Lo WHO protocol. Electropherotypes was observed in 10% po’ yacrylamide slab gel at 100V for 4 hour and staining with silver nitrate. During ihe 6-month study stool samples collected from 85 infants <5 years old, from RSU Kola Yogyakarra. ELISA carried out on 84 samples, 44 (5 2%) of positive rotavìrus. Genotyping can be performed on 43 samples, (il was found in 34 (79%) samples, G2 in 9 (21%). PAGE of 43 samples showed Cl has a long pattern, while G2 has short patterns. When compared with strains from other regions in Indonesia, 4 different patterns was found. Can be concluded strains G 1 and 09 has a long pattern, while the 02 and 03 has a short pattern.

Kata Kunci : Rotavirus,G,type,PAGE,Elektroferotipe,Rotavirus, G-type, PAGE, Electropherotype