Polyclonal antisera were raised against isolates of bean
common mosaic virus (BCMV) and bean common mosaic necrosis
virus (BCMNV) using conventional serological methods.
Infected tissues containing, respectively, 22 recognized BCMV
and BCMNV isolates were tested against the two antisera by
antigen-coated plate (ACP) ELISA and double antibody sandwich
(DAS) ELISA. Results indicated that each immunoglobulin was
virus-specific by DAS-ELISA, providing clear distinction
between BCMV and BCMNV.
A reverse transcription, polymerase chain reaction (RT-PCR)-based assay in combination with restriction endonuclease
analyses, was developed for molecular detection of BCMV, BCMNV
and their pathogroups. Specific detection of the two viruses
was accomplished by constructing two virus-specific primer
pairs that amplified a PCR product specific for each virus.
Distinction of two BCMNV pathogroups (PG-III and PG-VI) was
achieved by restriction enzyme XbaI digestion of BCMNV PCR
products. However, none of the tested restriction enzymes
clearly differentiated the five recognized BCMV pathogroups.
A primer pair Dts/Uny15 specific for BCMV pathogroup V was
also developed. By its RT-PCR application, four BCMV-PG-V
isolates were differentiated from the other known variants of
BCMV pathogroup I, II, IV and VII. Thus, by a combination of
RT-PCR and restriction enzyme analyses, it was possible to
differentiate both viruses, and two pathogroups of BCMNV, and
one pathogroup of BCMV. / Graduation date: 1996
| Identifer | oai:union.ndltd.org:ORGSU/oai:ir.library.oregonstate.edu:1957/34991 |
| Date | 30 June 1995 |
| Creators | Xu, Ling, 1963- |
| Contributors | Hampton, Richard O. |
| Source Sets | Oregon State University |
| Language | en_US |
| Detected Language | English |
| Type | Thesis/Dissertation |
Page generated in 0.0015 seconds