Deteksi penyakit TSV (Taura Syndrome Virus) secara PCR udang vannamei (Litopenaeus vannamei) dengan berbagai ekstraksi, suhu dan waktu penyimpanan

Abstract

The shrimp farming industries have been adversely affected by epizootics due to viral pathogens, especially Taura Syndrome Virus (TSV The TSV has been the causative agent of economically disastrous epizootics in Penaeus vannamei, causing mass mortalities of 40% - 90% in affected post larval and juvenile population. The current diagnostic and detection methods for TSV included clinical signs, gross pathology, in situ hybridization, and PCR. Three RNA extraction methods, i.e. RNA lysis buffer with Guanidine- HCL, RNA lysis buffer with beta-mercaptoethanol dan phenolchloroform) were used to evaluate the vannamei shrimp infected with TSV The study showed that RNA extracted using the RNA lysis buffer with beta-mercaptoethanol, constantly had the highest yield as measured (quality and quantity) using a geneQuant spectrophotometer at period of 3 mo11th in -20 OC storeage, except the one extracted by phenol - chloroform extraction had the highest yield quantity at -80 'C. Each of 3 (three) extraction methods yielded sufficient RNA for positive results in a RT PCR for TSV at period of 1 mo;zth,and 2 months, in both tempaature storage (-20°C and -800C), but at period of3 months, only the phenol- chloroform extraction give positive result after it was stored at -20°C and -80 °C.