PURIFICATION OF LILY SYMPTOMLESS CARLAVIRUS AND DETECTION OF THE VIRUS IN LILIES

Lily symptomless carlavirus (LSV) was purified from infected lilies (L ilium longiflorum) by employing Nonidet P-40, butanol, and urea during clarification followed by equilibrium centrifugation in cesium chlori de. Virus yields averaged 8.5 mg per 100 g of tissues using an extinct ion coefficient of 2.5. Two of the four rabbits injected three times e ach with 0.25 mg purified LSV had antiserum dilution endpoints of 10(- 5.4) by enzyme-linked immunosorbent assay (ELISA) or tissue-blot immun oassay (TBIA) at the first bleeding; whereas the other two rabbits had serum dilution endpoint of 10(-6). An indirect immunological procedur e was used to detect LSV antigens in tissue blots on nitrocellulose me mbranes. A comparison of ELISA and dot-blot immunoassay (DBIA) showed that DBIA was 16 to 32 times as sensitive as ELISA in detection of LSV . LSV was detected in bulb scales by TBIA in samples in which sap extr acts from the same scale pieces were negative in both DBIA and ELISA.