Studi Komparasi Metode Diagnosa Konvensional dan Pengembangan One Step Reverse Transcriptase Polymerase Chain Reaction untuk Deteksi Rabies
YUDIANINGTYAS, Dini Wahyu, Heru Susetya
2010 | Tesis |Penyakit rabies telah menyebar di wilayah timur Indonesia, sampai saat ini hanya dua propinsi yang masih bebas dari rabies di wilayah timur Indonesia (Propinsi Papua dan Irian Jaya Barat). Metode uji konvensional diagnosa rabies di Indonesia dilakukan secara pararel dengan urutan pewarnaan Sellers, Fluorescent Antibody Test (FAT) dan uji inokulasi pada mencit. Uji pewarnaan Seller memiliki beberapa kelebihan, sehingga bisa diaplikasikan di laboratorium tipe B maupun C khususnya di wilayah endemis rabies.Namun kelayakan uji pewarnaan Sellers sudah mulai diragukan oleh dunia internasional sebagai salah satu sarana peneguh diagnosa rabies. Pengembangan teknik diagnosa molekuler masih terus dilakukan dengan Reverse Transcriptase Polymerase Chain Reaction (RT-PCR). Penelitian ini bertujuan untuk mengevaluasi kesesuaian metode diagnosa rabies pewarnaan Seller dan RT-PCR dengan FAT serta optimasi onestep RT-PCR untuk diagnosa rabies. Pengujian dilakukan terhadap 153 sampel rabies yang ada di laboratorium virologi Balai Besar Veteriner Maros. Hasil analisa dengan uji Kappa menunjukkan bahwa kesesuaian pewarnaan Sellers terhadap FAT berkisar 0,75 – 0,80 sedangkan RT-PCR terhadap FAT berkisar 0,78 – 0,89. Hasil penelitian menunjukkan kesesuaian yang tinggi baik dari pewarnaan Sellers dan RT-PCR terhadap FAT serta tidak adanya kejadian negatif palsu dari kedua metode. Dari hasil penelitian dapat disimpulkan bahwa pewarnaan Sellers masih layak dipakai untuk diagnosa rabies di wilayah Indonesia timur sedangkan RT-PCR dapat dipertimbangkan sebagai sarana diagnosa rutin rabies. Kata kunci : virus rabies, Seller, FAT, RT-PCR, Kappa
Rabies disease has been spread in East Indonesia. To date only two provinces in East Indonesia are free from rabies disease (Papua Province and West Papua). Rabies conventional diagnostic’s method are applied as a serial using Sellers’s staining Fluorescent Antibody Test (FAT) and mouse inoculation test. Sellers’s staining has some advantages and can be apllied by B and C type laboratories, particulary at endemic areas. However, Seller’s staining method has been questioned by international community for its accuracy as a rabies diagnostic tool. Development of molecular based technique still on continuing process using Reverse Transcriptase Polymerase Chain Reaction (RT-PCR). Aim of this study are to evaluate and describe out agreement value of Sellers staining and RT-PCR compared to FAT and optimation of one-step RT-PCR for rabies detection. Out of 130 rabies samples tested at virology and biotecnology laboratories of Disease Investigation Centre Maros. Results of Kappa showed that agreement of Sellers’s staining to FAT were 0,75–0,80 and agreement value of one-step RT-PCR to FAT were 0,86–0,89 also there was no false negative result found. These results indicated high agreements from both Sellers’s staining and one-step RT-PCR. These findings suggest that both techniques, Sellers’s staining and one-step RT-PCR could be considered as tools for rabies eradication program. Further studies are required to assess the sensitivity and specificity of the rabies diagnostic tests using the gold standard. Kata kunci : virus rabies, Seller, FAT, RT-PCR, Kappa
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