Canadian Forest Service Publications

Use of monoclonal antibody against poly IS: poly CS for detecting mycoviruses and potential applications to potato spindle tuber viroid and animal reoviruses. 1987. Benhamou, N.; Parent, J.G.; Garzon, S.; Asselin, A.; Ouellette, G.B.; Joly, J.R. Canadian Journal of Plant Pathology 9: 106-114.

Year: 1987

Issued by: Laurentian Forestry Centre

Catalog ID: 14302

Language: English

Availability: PDF (request by e-mail)

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Seven hybridoma cell lines, produced by fusing myeloma cells and splenocytes from BALB/c mice immunized with poly [I]: poly [C] complexed to methylated bovine serum albumin (MBSA), were selected for anti- dsRNA monoclonal antibodies (MAbs) by ELISA. Two cell lines 24-4D4 and 24-3A8, reacted positively with synthetic poly[I]: poly [C] and poly [A]: poly [U], whereas no reaction was observed with synthetic or naturally occurring ssRNA, and DNA. Sensitivity of 24-3A8 MAb was assessed by detecting amounts as low as 5 to 20 ng of potato spindle tyber viroid (PSTV) in infected potato leaf extract. In a screening program to detect mycoviruses by dot-immunobinding procedure, 64% of the 100 Ascocalyx abietina isolates tested were found positive for dsRNA. The presence of mycoviruses did not appear to be related to race classification. The nonaggressive Q412 strain of Ophiostoma ulmi was positive for dsRNA, whereas no reaction was noted with the aggressive 695 strain. Ultrastructural investigations of fungal material revealed a very low degree of virus infection. The potential applicability of 24-3A8 MAb to recognize ultrastructurally dsRNA was tested by an immunogold procedure on reovirus-infected avian cells. No gold labeling was noted with ssRNA virus-infected animals cells. The absence of labeling of the dsRNA reovirus involved in the cytoplasmic polyhedrosis of insects suggested the importance of the conformational structure for antigenicity.