In Ethiopia five species of tsetse flies were recorded from different parts of therncountry. Glossina pallidipes is one of the five species which cause a fatal disease tornhuman and animal. Sterile Insect Technique (SIT) as a component for tsetse flyrncontrol and eradication programme is one of the successful techniques in somerninsect pest eradication programmes. However, the presence of salivary glandrnhyperplasia virus affects the application of SIT technique in tsetsecontrol as itrnlimits the mass rearing activity. It is widely accepted that tsetse plays host tornsalivary gland hyperplasia virus. This study mainly focused on virus free linernestablishment and the antiviral drug treatment using more than 1800 G.pallidipesrnTororo and 3072 G. pallidipes Arbaminch strains. Prior to the large scalernexperiments, two preliminary surveys were done on virus free line establishmentrnand antiviral drug treatments. All experiments were analyzed using PCR,rndissection and QPCR methods. The preliminary survey on the investigation ofrnthe virus free line by dissection showed that 4.45% male flies and 3.09% of femalernflies were positive for salivary gland hyperplasia, whereas in PCR analysis 95% of thernflies were positive for SGHV. In the virus free line establishment a total of 920 femalernflies were taken by randomly and 13.04% were selected by PCR as positive or negativernflies for salivary gland hyperplasia and two out of 120 female flies died. Amongrn120 flies, 26.7% male and 15% female including the dead flies were negative andrn35.8% female and 22.5% male flies were positive by PCR. All screened flies wererngrouped into four groups as Virus free line I, II, III and IV. In the preliminaryrnsurvey of the antiviral drug, female flies treated with valacyclovir treated fliesrnproduced more pupae than acyclovir treated flies. In the main experiment of thernantiviral drugs valacyclovir treated flies showed no significant difference P>.072.rnBut, acyclovir and control showed significant difference P