One-Step Reverse-Transcription Digital PCR for Reliable Quantification of Different Pepino Mosaic Virus Genotypes

Plants
Nataša MehleDavid Dobnik

Abstract

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 ...Continue Reading

References

Aug 4, 1999·Proceedings of the National Academy of Sciences of the United States of America·B Vogelstein, K W Kinzler
Oct 12, 2002·Archives of Virology·J M AguilarM A Aranda
Sep 1, 2004·BioTechniques·Peter M Vallone, John M Butler
Aug 1, 2009·Journal of Virological Methods·Ion Gutiérrez-AguirreMaja Ravnikar
Nov 30, 2011·Analytical Chemistry·Leonardo B PinheiroKerry R Emslie
Jun 13, 2012·Microbiology and Molecular Biology Reviews : MMBR·Esteban DomingoCelia Perales
Mar 9, 2013·PloS One·Alexandra S WhaleJim F Huggett
Dec 18, 2013·Applied and Environmental Microbiology·N MehleM Ravnikar
Jan 7, 2014·Journal of Virology·Manuel G Moreno-PérezFernando García-Arenal
Dec 19, 2016·Biomolecular Detection and Quantification·J MadicM E Droniou
Sep 19, 2017·Analytical and Bioanalytical Chemistry·Alexandra Bogožalec KoširJana Žel
Feb 19, 2019·Analytical Chemistry·Kerry R EmslieDaniel G Burke
Jan 1, 2000·Plant Disease·R A A van der VlugtD-E Lesemann
May 28, 2019·Food Chemistry·Alexandra Bogožalec KoširMojca Milavec

❮ Previous
Next ❯

Methods Mentioned

BETA
PCR
genotyping
reverse-transcription dPCR
chips

Software Mentioned

QuantaSoft
Naica
Cristal Miner
SDS

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.