Molecular detection of nine rice viruses by a reverse-transcription loop-mediated isothermal amplification assay.
A reverse-transcription loop-mediated isothermal amplification (RT-LAMP) assay was established for the detection of nine viruses from infected rice plants, including rice black-streaked dwarf virus (RBSDV), rice dwarf virus (RDV), rice gall dwarf virus (RGDV), rice ragged stunt virus (RRSV), rice transitory yellowing virus (RTYV), rice stripe virus (RSV), rice grassy stunt virus (RGSV), rice tungro spherical virus (RTSV), and rice tungro bacilliform virus (RTBV). Virus-specific primer sets were designed from the genome sequences of these viruses. By the combination of RNA rapid extraction and RT-LAMP, these nine viruses could be detected within 2h from infected rice plants. The sensitivities of the assays were either higher than (for RSV, RTBV, and RTYV) or similar (for RDV) to those of one-step RT-PCR. Furthermore, RTBV and RTSV were detected not only in infected rice plants but also in viruliferous insect vectors. The RT-LAMP assays may facilitate studies on rice disease epidemiology, outbreak surveillance, and molecular pathology.
Le
Dung Tien
DT
Research Team for Vector-Borne Diseases, National Agricultural Research Center, Tsukuba, Ibaraki, Japan.
Netsu
Osamu
O
Uehara-Ichiki
Tamaki
T
Shimizu
Takumi
T
Choi
Il-Ryong
IR
Omura
Toshihiro
T
Sasaya
Takahide
T
D000383:Agriculture; D000818:Animals; D001483:Base Sequence; D017124:Conserved Sequence; D017931:DNA Primers; D004279:DNA, Viral / Q000235:genetics; D007303:Insect Vectors / Q000821:virology; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D012275:Oryza / Q000821:virology*; D018515:Plant Leaves / Q000821:virology; D010942:Plant Viruses / Q000235:genetics / Q000302:isolation & purification*; D048348:Reverse Transcription; D012680:Sensitivity and Specificity; D015678:Viral Structural Proteins / Q000235:genetics
D017931:DNA Primers; D004279:DNA, Viral; D015678:Viral Structural Proteins