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Sequence-modified primers for the differential RT-PCR detection of Andean potato latent and Andean potato mild mosaic virus in quarantine tests

GND
1225899559
Affiliation
Julius Kühn-Institute (JKI), Institute for Epidemiology and Pathogen Diagnostics, Germany
Koenig, Renate;
GND
1058967991
Affiliation
Julius Kühn-Institute (JKI), Institute for Epidemiology and Pathogen Diagnostics, Germany
Ziebell, Heiko

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.

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