Sequence-modified primers for the differential RT-PCR detection of Andean potato latent and Andean potato mild mosaic viruses in quarantine tests.
To enable the differential PCR detection of Andean potato latent virus (APLV) and Andean potato mild mosaic virus (APMMV) strains, sense primers were designed that correspond to regions directly upstream of the coat protein genes. Their differentiating power was increased by A->C or T->C replacements in their 3'-terminal parts. Together with the broad-specificity antisense primer EM3, primer AL-a-mod3C detected all APLV strains tested, but none of the APMMV strains. Primer AM-a-mod4C yielded PCR products with all APMMV preparations, but also with some APLV preparations. Sequence analysis revealed that this was not due to a lack of primer specificity, but to the sensitive detection of contaminating APMMV in some of our APLV preparations.
c/o Julius Kühn Institute, Institute for Epidemiology and Pathogen Diagnostics, Messeweg 11, 38104, Braunschweig, Germany, renate.koenig@jki.bund.de.
Renate
R
Koenig
Heiko
H
Ziebell
D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics*; D009029:Mosaic Viruses / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification*; D010935:Plant Diseases / Q000821:virology*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D011198:Solanum tuberosum / Q000821:virology*
D017931:DNA Primers