title: The development and application of a Dot-ELISA assay for diagnosis of southern rice black-streaked dwarf disease in the field.
abstract: Outbreaks of the southern rice black-streaked dwarf virus (SRBSDV) have caused significant crop losses in southern China in recent years, especially in 2010. There are no effective, quick and practicable methods for the diagnosis of rice dwarf disease that can be used in the field. Traditional reverse transcription-polymerase chain reaction (RT-PCR) methodology is accurate but requires expensive reagents and instruments, as well as complex procedures that limit its applicability for field tests. To develop a sensitive and reliable assay for routine laboratory diagnosis, a rapid dot enzyme-linked immunosorbent assay (dot-ELISA) method was developed for testing rice plants infected by SRBSDV. Based on anti-SRBSDV rabbit antiserum, this new dot-ELISA was highly reliable, sensitive and specific toward SRBSDV. The accuracy of two blotting media, polyvinylidene fluoride membrane (PVDF membrane) and nitrocellulose filter membrane (NC membrane), was compared. In order to facilitate the on-site diagnosis, three county laboratories were established in Shidian (Yunnan province), Jianghua (Hunan Province) and Libo (Guizhou province). Suspected rice cases from Shidian, Yuanjiang and Malipo in Yunnan province were tested and some determined to be positive for SRBSDV by the dot-ELISA and confirmed by the One Step RT-PCR method. To date, hundreds of suspected rice samples collected from 61 districts in southwestern China have been tested, among which 55 districts were found to have rice crops infected by SRBSDV. Furthermore, the test results in the county laboratories showed that Libo, Dehong (suspected samples were sent to Shidian) and Jianghua were experiencing a current SRBSDV outbreak.
authors.affiliation: State Key Laboratory Breeding Base of Green Pesticide and Agricultural Bioengineering/ Key Laboratory of Green Pesticide and Agricultural Bioengineering, Ministry of Education, Guizhou University, Guiyang, 550025, PR China. gs.zcwang@mail.gzu.edu.cn
authors.forename: Zhenchao
authors.initials: Z
authors.lastname: Wang
authors.forename: Dandan
authors.initials: D
authors.lastname: Yu
authors.forename: Xiangyang
authors.initials: X
authors.lastname: Li
authors.forename: Mengjiao
authors.initials: M
authors.lastname: Zeng
authors.forename: Zhuo
authors.initials: Z
authors.lastname: Chen
authors.forename: Liang
authors.initials: L
authors.lastname: Bi
authors.forename: Jiaju
authors.initials: J
authors.lastname: Liu
authors.forename: Linhong
authors.initials: L
authors.lastname: Jin
authors.forename: Deyu
authors.initials: D
authors.lastname: Hu
authors.forename: Song
authors.initials: S
authors.lastname: Yang
authors.forename: Baoan
authors.initials: B
authors.lastname: Song
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology; D000918:Antibody Specificity; D001483:Base Sequence; D015153:Blotting, Western / Q000379:methods; D002681:China; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D016679:Genome, Viral; D008969:Molecular Sequence Data; D012275:Oryza / Q000821:virology*; D010935:Plant Diseases / Q000821:virology*; D012367:RNA, Viral / Q000032:analysis*; D011817:Rabbits; D012087:Reoviridae / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D012689:Sequence Homology, Nucleic Acid; D012756:Sheep; D013045:Species Specificity
chemical_list: D000914:Antibodies, Viral; D012367:RNA, Viral