title: Detection of tomato yellow leaf curl Thailand virus by PCR without DNA extraction.
abstract: We report the simple and rapid method for detection of tomato yellow leaf curl Thailand virus (TYLCTHV) based on the direct capture of virus particles to the surface of a polymerase chain reaction (PCR) tube. This method allowed PCR without the time-consuming procedures of DNA extraction from infected plant tissue. A small amount of tomato tissue (approximately 10 mg) was ground in extraction buffer to release viruses from plant tissues. The constituents of the plant extract that might inhibit PCR activity were discarded by washing the tube with PBST buffer before adding the PCR mixture to the tube. This method was used for detection of TYLCTHV with plant sap solution diluted up to 1:20,000 and was more sensitive than an enzyme-linked immunosorbent assay (ELISA) method. In addition, this method can be used for detection of TYLCTHV in viruliferous whiteflies. The PCR tubes with captured TYLCTHV could be used for PCR, after storage at 4 degrees C for 4 wk. The method presented here was used for detection of begomoviruses in cucurbit and pepper. In addition, this method was effectively used to detect papaya ringspot virus in papaya and zucchini yellow mosaic virus in cucumber by reverse transcriptase (RT)-PCR.
authors.affiliation: Center for Agricultural Biotechnology, Kasetsart University, Kamphaengsaen Campus, NP 73140 Thailand.
authors.forename: Supaporn
authors.initials: S
authors.lastname: Ieamkhang
authors.forename: Lumpueng
authors.initials: L
authors.lastname: Riangwong
authors.forename: Orawan
authors.initials: O
authors.lastname: Chatchawankanphanich
mesh_terms: D000818:Animals; D053839:Begomovirus / Q000235:genetics* / Q000302:isolation & purification*; D004279:DNA, Viral / Q000235:genetics*; D018551:Solanum lycopersicum / Q000821:virology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
chemical_list: D004279:DNA, Viral