Comparison of the Passive Haemagglutination and Bentonite Flocculation Tests for Serological Work With Plant Viruses

The passive haemagglutination and bentonite flocculation tests were compared for detecting plant viruses, using antibody-sensitized tanned red cells or bentonite particles. Parsnip yellow fleck, raspberry ringspot, turnip yellow mosaic, narcissus mosaic, potato X and tobacco rattle viruses were detected by both tests in purified preparations and, where studied, in crude plant extracts. The highest antigen titres were obtained only when the red cells or bentonite particles were sensitized with optimal amounts of antibody; these had to be found experimentally because they differed between antisera and between the two tests. In comparative experiments the bentonite flocculation and passive haemagglutination tests were respectively about 2 to 5 and IOO to 125 times more sensitive than tube-precipitin tests for detecting the elongated viruses and about IO to 2o and 5oo to 6oo times more sensitive for detecting the isometric viruses. The minimum concentrations of virus detected were 1.8 to 2.6/,g./ml. with the bentonite flocculation test and o'o4 to o'24/,g./ml, with the passive haemagglutination test. Red cells treated with formalin before they were tanned and sensitized with antibody could be preserved at -14 ° for up to 8 weeks without loss of activity. In experiments with turnip yellow mosaic virus, extracts from healthy plants of eight species tested had little or no effect on either test but concentrated extracts from Chenopodium amaranticolor and Spinacia oleracea plants caused non-specific agglutination of unformalinized red cells, and dilute extracts from Petunia hybrida plants decreased eightfold the sensitivity of the passive haemagglutination test. These effects did not occur with formalinized red cells. Antibody-sensitized red cells were used in haemagglutination-inhibition tests to determine antiserum titres; antisera to turnip yellow mosaic and raspberry ringspot viruses gave titres 32 times higher than those obtained in tubeprecipitin tests.