Purification of Lily Symptomless Carlavirus and Detection of the Virus in Lilies. H. T. Hsu, Microbiologist; Department of Agriculture, Agricultural Research Service, Floral and Nursery Plants Research Unit, Beltsville Agricultural Research Center, Beltsville, Md. 20705-2350. J. Y. Kim, Visiting Scientist, and R. H. Lawson, Research Plant Pathologist, U.S. Department of Agriculture, Agricultural Research Service, Floral and Nursery Plants Research Unit, Beltsville Agricultural Research Center, Beltsville, Md. 20705-2350. Plant Dis. 79:912-916. Accepted for publication 1 May 1995. This article is in the public domain and not copyrightable. It may be freely reprinted with customary crediting of the source. The American Phytopathological Society, 1995. DOI: 10.1094/PD-79-0912.

Lily symptomless carlavirus (LSV) was purified from infected lilies (Lilium longiflorum) by employing Nonidet P-40, butanol, and urea during clarification followed by equilibrium centrifugation in cesium chloride. Virus yields averaged 8.5 mg per 100 g of tissues using an extinction coefficient of 2.5. Two of the four rabbits injected three times each with 0.25 mg purified LSV had antiserum dilution endpoints of 10-5,4 by enzyme-linked immunosorbent assay (ELISA) or tissue-blot immunoassay (TBIA) at the first bleeding; whereas the other two rabbits had serum dilution endpoint of 10-6. An indirect immunological procedure was used to detect LSV antigens in tissue blots on nitrocellulose membranes. 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 extracts from the same scale pieces were negative in both DBIA and ELISA

Keyword(s): disease indexing method, immunohistochemical detection, immunological assay, serodiagnosis, virus localization, virus purification