Methodology for antibody preparation and detection of southern rice black-streaked dwarf virus.
Southern rice black-streaked dwarf virus (SRBSDV) is a member of the genus Fijivirus of the family Reoviridae that causes a devastating disease in rice. Seven polypeptide fragments of the Putative uncharacterized protein (Pup) and the P10 proteins of SRBSDV were designed, synthesized, and used to immunize rabbits. Titers of polyclonal antibodies against the seven peptides were examined using indirect enzyme-linked immunosorbent assay (ELISA), and their specificities were investigated using western blotting. Indirect dot-immunobinding assay (DIBA) was also carried out at different dilutions against an antigen (rice extract). Antibody-1, which had the highest selectivity and titer, was then used to examine rice samples suspected of being infected with SRBSDV that were collected for over two years in different areas of China, using DIBA. Our results indicate that antibody-1 has the advantages of reliability, high sensitivity, and high specificity. Use of this antibody can help facilitate identification of the virus and its distribution in rice-growing areas where it causes significant problems.
State Key Laboratory Breeding Base of Green Pesticide and Agricultural Bioengineering, Key Laboratory of Green Pesticide and Agricultural Bioengineering, Guizhou University, Ministry of Education, Guiyang 550025, People's Republic of China.
Zhuo
Z
Chen
Chengjun
C
Yin
Jiaju
J
Liu
Mengjiao
M
Zeng
Zhenchao
Z
Wang
Dandan
D
Yu
Liang
L
Bi
Linhong
L
Jin
Song
S
Yang
Baoan
B
Song
D000818:Animals; D000914:Antibodies, Viral / Q000096:biosynthesis* / Q000276:immunology; D000918:Antibody Specificity; D000956:Antigens, Viral / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D015151:Immunoblotting / Q000379:methods; D008856:Microscopy, Fluorescence; D016253:Microscopy, Immunoelectron; D012275:Oryza / Q000821:virology*; D010935:Plant Diseases / Q000821:virology; D011817:Rabbits; D012087:Reoviridae / Q000276:immunology* / Q000302:isolation & purification*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D012680:Sensitivity and Specificity; D014764:Viral Proteins / Q000737:chemistry / Q000276:immunology
D000914:Antibodies, Viral; D000956:Antigens, Viral; D014764:Viral Proteins