Reverse transcription loop-mediated isothermal amplification of RNA for sensitive and rapid detection of southern rice black-streaked dwarf virus.
Southern rice black-streaked dwarf virus (SRBSDV) causes one of the most serious viral diseases of rice in Southeast Asia. A reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for detection of SRBSDV using total RNA extracted from rice tissues and the insect pest, white-backed planthopper. The assay was based on a set of four primers matching a total of six sequences in the S9 region of SRBSDV genome. Presence of the virus could be detected in RT-LAMP reactions containing 1.2×10(-6)μg of a total RNA extract, which was ten times more sensitive than a classical RT-PCR assay. The SRBSDV could be distinguished from the closely related rice black-streaked dwarf virus (RBSDV) by this method, indicating a high degree of specificity. This simple and sensitive RT-LAMP assay shows potential for detection of SRBSDV in field samples of hosts or vectors at a relatively low cost.
College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, China.
Tong
T
Zhou
Linlin
L
Du
Yongjian
Y
Fan
Yijun
Y
Zhou
D001210:Asia, Southeastern; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D012275:Oryza / Q000821:virology*; D010935:Plant Diseases / Q000821:virology*; D012367:RNA, Viral / Q000032:analysis* / Q000302:isolation & purification; D012087:Reoviridae / Q000235:genetics* / Q000302:isolation & purification*; D048348:Reverse Transcription; D012680:Sensitivity and Specificity; D013997:Time Factors
D012367:RNA, Viral