20.500.12556/DiRROS-19509 One-step reverse-transcription digital PCR for reliable quantification of different pepino mosaic virus genotypes In recent years, pepino mosaic virus (PepMV) has rapidly evolved from an emerging virus to an endemic pathogen, as it causes significant loses to tomato crops worldwide. At present, the main control strategy for prevention of PepMV disease in tomato production remains based on strict hygiene measures. To prevent damage caused by PepMV, cross-protection is used in some countries. Reliable characterisation, detection and quantification of the pathogen are vital for disease control. At present, reverse-transcription real-time quantitative polymerase chain reaction (RT-qPCR) is generally used for this purpose. However, quantitative use of RT-qPCR is linked to standardised reference materials, which are not available for PepMV. In addition, many factors can influence RT-qPCR efficiencies and lead to lower accuracy of the quantification. In this study, well-characterised PepMV-genotype-specific RT-qPCR assays were transferred to two digital PCR (dPCR) platforms. dPCR-based assays allow absolute quantification without the need for standard curves, and due to the binary nature of the reaction, dPCR also overcomes many of the other drawbacks of RT-qPCR. We have shown that these newly developed and validated PepMV-genotype-specific dPCR assays are suitable candidates for higher-order methods for quantification of PepMV RNA, as they show lower measurement variability, with sensitivity and specificity comparable to RT-qPCR. digital PCR pepino mosaic virus quantification genotype specific true false true Angleški jezik Ni določen Neznano 2024-07-19 03:42:01 2024-07-19 13:59:54 2024-07-20 04:03:07 0000-00-00 00:00:00 2020 0 0 Nasl. z nasl. zaslona; Opis vira z dne 10. 3. 2020; str. 1-13 no. 3 Vol. 9 2020 0000-00-00 Zaloznikova Objavljeno NiDoloceno 0000-00-00 0000-00-00 2020-03-04 577.2 2223-7747 10.3390/plants9030326 5319247 https://www.mdpi.com/2223-7747/9/3/326 1 5418e387-455f-11ef-b487-001a4af901a5 https://dirros.openscience.si/Dokument.php?lang=slv&id=26917 RAZ_Mehle_Natasa_2020.pdf RAZ_Mehle_Natasa_2020.pdf 1 DB407068B34B85801FF2637758065E9D 2b4670f4ebd425a3d1237802e43fdea7b85cccca8d7be57ea612d849e235815d dddd5362-45b5-11ef-b487-001a4af901a5 https://dirros.openscience.si/Dokument.php?lang=slv&id=27025 Nacionalni inštitut za biologijo 0 0 0