【Objective】The present study is an attempt to produce Sugarcane yellow leaf virus (SCYLV) free plantlets from commercial sugarcane varieties， UP 01235 and UP 0097， using meristem tip culture technique. 【Method】Plantlets were produced using meristem (different size) as explant， collected from virus infected and healthy field grown sugarcane plants. RT-PCR assay of regenerated plantlets was carried out to check the presence of virus at monthly interval up to six month stage using specific primer pair (SCYLV-615F：5′-ATGAATACGGGCGCTAACCGYYCAC-3′ and SCYLV-615R：5′-GTGTTGGGGRAGCGTCGCYTACC-3′) obtained from Sugarcane yellow leaf virus. 【Result】The plantlets of both varieties， regenerated from the meristems measuring up to 1.5 mm， did not showed the presence of virus during any growth stage under glasshouse conditions， this indicated the absolute elimination of SCYLV. However， the meristems measuring more than 1.5 mm size showed presence of the SCYLV in PCR assay.【Conclusion】More than 200 plantlets， obtained from 1.0~1.5 mm meristem tip culture were propagated in greenhouse showed absence of SCYLV during all growth stages.