|
Abstract The aim of this assay is to investigate on the TaqMan Real-time PCR method for phytoplasma general detection which has the superiorities in saving time and preventing contamination. One developed TaqMan Real-time PCR assay for general detection of phytoplasma was evaluated in comparison to conventional nested-PCR method with the aim to assess its potential for research and routine applications. In this study, specificity and sensitivity of the Real-time PCR was evaluated using 12 phytoplasma infected and 9 healthy plants DNA. All the phytoplasma got high fluorescence signals, healthy plant DNA and blank control got negative results. The TaqMan Real-time PCR procedure had high test specificity and higher sensitivity 10 times than that of the nested-PCR. The TaqMan Real-time PCR had the simplest and fastest testing process, especially decreased the risk of PCR contamination. Results inclicate that has a high potential for automation, and seems to represent the currently most suitable method for large-scale phytoplasma general detection.
|
|
Received: 08 December 2010
|
|
|
|
