Distinguishing Between Two Closely Related Strains of Xylella fastidiosa
||International Journal of Biotech Trends and Technology (IJBTT)|
|© 2014 by IJBTT Journal|
|Volume - 4 Issue - 4
|Year of Publication : 2014|
|Authors : Arinder K. Arora, Ravi Durvasula, Thomas A. Miller|
Arinder K. Arora, Ravi Durvasula, Thomas A. Miller "Distinguishing Between Two Closely Related Strains of Xylella fastidiosa", International Journal of Biotech Trends and Technology (IJBTT), V4(4):8-13 Oct - Dec 2014, Published by Seventh Sense Research Group.
A real time PCR assay (rt-PCR), using Taqman minor groove binder (MGB) probes was developed to quantify and differentiate two strains of Xylella fastidiosa, Pierce’s Disease (PD) strain and Almond Leaf Scorch (ALS) strain. Two isolates, Temecula and Dixon, were used as references for the PD strain and the ALS strain, respectively. Primers and probes based on the gyraseB sequence were able to distinguish between both the isolates, and DNA concentrations as low as 1.01×10-5 ng/µl (3 copy number/µl) were detected. MGB probes were also able to distinguish between the Temecula and Dixon isolate DNA from a mixture of the two DNA’s. Whereas the Temecula specific MGB probe showed 106–fold resolution, the Dixon specific probe showed only 10-fold resolution. We have developed a reliable, sensitive and specific rt-PCR assay using MGB probes to quantify and distinguish DNA of the two strains of X. fastidiosa.
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Real time-PCR, Xylella fastidiosa, strains, PD, ALS, Temecula, Dixon.