Detecting Rice stripe virus (RSV) in the small brown planthopper (Laodelphax striatellus) with high specificity by RT-PCR.
Rice stripe disease, caused by Rice stripe virus (RSV), may lead to severe or even crippling losses in many rice-cultured countries and regions. As the most important vector of RSV, the small brown planthopper (SBPH) (Laodelphax striatellus) is largely responsible for the epidemic phase of the disease. Therefore, a rapid identification of RSV in the SBPH is of a great need for disease forecasting. A reverse transcription polymerase chain reaction (RT-PCR) assay is described to amplify a RSV gene in individual L. striatellus. By using primers matched to the viral RNA dependent RNA polymerase gene in RNA1, a 445 bp product was detected in viruliferous SBPHs. Meanwhile, the PCR products produced by the SBPH actin primers constructed across the boundary of an intron and an exon were used as RNA specific positive control for each stage of the experiment to ensure the validity of the negative results. Duplex RT-PCR conditions were established for the simultaneous detection of RSV and actin. This approach can be used for the early detection of RSV in L. striatellus and the subsequent rice stripe disease forecasting.
Lijun
Cai
C
State Key Laboratory of Genetic Engineering, Department of Life Science, Institute of Genetics, School of Life Science, Fudan University, Handan Road 220, Shanghai 200433, China.
Xizhi
Ma
M
Lin
Kang
K
Kejing
Deng
D
Shouyuan
Zhao
Z
Changben
Li
L
D000818:Animals; D017931:DNA Primers; D007313:Insecta / Q000201:enzymology / Q000235:genetics / Q000821:virology*; D012275:Oryza / Q000821:virology*; D010942:Plant Viruses / Q000235:genetics / Q000302:isolation & purification; D012367:RNA, Viral / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D029364:Tenuivirus / Q000235:genetics / Q000302:isolation & purification*
D017931:DNA Primers; D012367:RNA, Viral