Detection of Tomato Yellow leaf Curl Thailand Virus by PCR Without DNA Extraction
Detection of Tomato Yellow leaf Curl Thailand Virus by PCR Without DNA Extraction
- Molecular Biotechnology, 31(3), p.233-238, 2005 .
We report the simple and rapid method detection of tomato yellow leaf curl Thailand virus (TYLCTHV)based on the direct capture of virus partic1es to the surface of a polymerase chain reaction (PCR)tube. This method allowed PCR without the time-consuming pracedures of DNA extraction fram infected plant tissue. A small amount of tomato was ground in extraction Buffer to release viruses fram plant tissues. The sconstituents 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 -withplant 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°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.
TOMATO YELLOW LEAF CTIRL THAILAND VIRUS
GEM.INIVIRUSES
BEGOMOVIRUSES
PCR
RT-PCR
DNA EXTRACTION.
We report the simple and rapid method detection of tomato yellow leaf curl Thailand virus (TYLCTHV)based on the direct capture of virus partic1es to the surface of a polymerase chain reaction (PCR)tube. This method allowed PCR without the time-consuming pracedures of DNA extraction fram infected plant tissue. A small amount of tomato was ground in extraction Buffer to release viruses fram plant tissues. The sconstituents 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 -withplant 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°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.
TOMATO YELLOW LEAF CTIRL THAILAND VIRUS
GEM.INIVIRUSES
BEGOMOVIRUSES
PCR
RT-PCR
DNA EXTRACTION.