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DETEKSI DAN IDENTIFIKASI ARBOVIRUS PENYEBAB DEMAM PADA PASIEN DI BADAN RUMAH SAKIT UMUM (BRSU) TABANAN, BALI

DETECTION AND IDENTIFICATION OF ARBOVIRUS AS THE CAUSE OF FEBRILE ILLNESS IN PATIENTS FROM TABANAN HOSPITAL, BALI

Undergraduate Theses from JBPTITBPP / 2017-09-28 10:27:58
Oleh : Andini Nurfatimah Karim - Nim: 10613048, S1 - Biology-SITH
Dibuat : 2017-09-28, dengan 1 file

Keyword : Arbovirus, chikungunya, demam, dengue, diagnosa virus, japanese encephalitis

Infeksi arbovirus (arthropod-borne virus) merupakan salah satu masalah kesehatan publik yang signifikan di dunia. Diagnosa klinis sulit dilakukan karena kebanyakan infeksi berlangsung tanpa disertai gejala, ataupun dengan gejala menyerupai infeksi virus/patogen lain. Bali, sebagai pusat pariwisata di Indonesia endemik virus dengue (DENV) dari famili Flaviviridae dan chikungunya (CHIKV) dari famili Alphaviridae. Penelitian ini bertujuan untuk mendeteksi dan mengidentifikasi arbovirus (Alphaviridae dan Flaviviridae) penyebab demam pada pasien di Badan Rumah Sakit Umum (BRSU) Tabanan, Bali, dengan metode molekuler dan serologi. Sampel berupa plasma darah, sejumlah 100 sampel akut dan 5 sampel konvalesens diambil dari pasien rawat inap dan rawat jalan di BRSU Tabanan, Bali, periode Januari-Maret 2017. Kriteria inklusi yaitu demam >38°C selama kurang dari 7 hari dengan leukosit <10.000 sel/μl. Metode molekuler dan tambahan isolasi virus dilakukan pada sampel akut, sementara uji serologi dilakukan pada sampel akut dan juga konvalesens. Metode molekuler diawali dengan RT-PCR tingkat famili Alphaviridae dan Flaviviridae yang dilanjutkan dengan RT-PCR tingkat spesifik dan/atau sekuensing DNA. Sekuensing gen envelop juga dilakukan untuk analisis filogenetik. Uji serologi dibagi menjadi ELISA IgM DEN, CHIK, dan JE (Japanese Encephalitis). Isolasi virus dilakukan dengan kultur sel vero81. Hasil RT-PCR mendeteksi 3 sampel positif Alphaviridae yang diidentifikasi sebagai CHIKV dan 32 sampel positif Flaviviridae yang diidentifikasi sebagai DENV (2 sampel DENV-1, 7 sampel DENV-2, 14 sampel DENV-3 dan 9 sampel DENV-4). Dari uji ELISA, ditemukan 1 sampel positif IgM DEN (positif RT-PCR) dan 1 sampel positif IgM JE (negatif RT-PCR). Hasil kultur sel vero81 menunjukkan cytopathic effect (CPE) pada 23/36 sampel positif RT-PCR dan ELISA. Melalui analisis filogenetik, diketahui ketiga sampel positif CHIKV termasuk genotipe Asia, 1 sampel positif masing-masing DENV-1 dan DENV-3 termasuk genotipe I, dan DENV-2 termasuk genotipe kosmopolitan. Dapat disimpulkan, arbovirus penyebab demam pada pasien di BRSU Tabanan Bali yaitu DENV (32%) dan JEV (1%) dari famili Flaviviridae dan CHIKV (1%) dari famili Alphaviridae.


Deskripsi Alternatif :

Infection of arbovirus (arthropod-borne virus) is one of the most significant public health issues in the world. Clinical diagnosis is challenging as most of the cases shown to be asymptomatic, or symptomatic but resemble other viral/pathogenic infection. Bali, as the center of world tourism in Indonesia is endemic for dengue virus (DENV) from Flaviviridae and chikungunya virus (CHIKV) from Alphaviridae. This research is aimed to detect and identify arbovirus (Alphaviridae and Flaviviridae) as the cause of febrile illness in patients from Tabanan hospital, Bali, using molecular and serological assay. Blood plasma samples, with total 100 acute and 5 convalescence samples were collected from in- and outpatients of Tabanan local hospital, Bali, on January-March 2017. Criteria of inclusion was fever >38°C for less than 7 days with leukocyte count <10.000 cells/μl. Molecular assay and virus isolation -as an addition- were conducted on acute samples, while serological assay was conducted on acute and convalescence samples. Molecular assay was started with RT-PCR in family level Alphaviridae and Flaviviridae and continued to RT-PCR specific level and/or DNA sequencing. Sequencing with envelope gene target was also conducted for phylogenetic analysis. Serological assay was divided into ELISA DEN, CHIK, and JE (Japanese Encephalitis) IgM. Virus isolation is conducted with vero81 cell culture. The RT-PCR results detected 3 positive samples of Alphaviridae and identified as CHIKV and 32 positive samples of Flaviviridae and identified as DENV (2 samples of DENV-1, 7 samples of DENV-2, 14 samples of DENV-3 and 9 samples of DENV-4). From ELISA test, found 1 positive sample of DEN IgM (positive RT-PCR) and 1 positive sample of JE IgM (negative RT-PCR). Vero81 cell culture results showed the cytopathic effect (CPE) on 23/36 positive samples of RT-PCR and ELISA. Through the phylogenetic analysis, 3 positive samples of CHIKV were belong the Asian genotype, 1 positive sample from each DENV-1 and DENV-3 was belong to genotype I, and from DENV-2 was belong to cosmopolitan genotype. It can be concluded, arbovirus which cause febrile illness in patients from Tabanan hospital, Bali were DENV (32%) and JEV (1%) from Flaviviridae and CHIKV (1%) from Alphaviridae.


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