Efficient methods for sample processing and cDNA synthesis by RT-PCR for the detection of grapevine viruses and viroids

Journal of Virological Methods
Ryoji Nakaune, Masaaki Nakano

Abstract

Template preparation is important in reverse-transcription polymerase chain reaction (RT-PCR)-based detection methods. A TissueLyser with tungsten carbide beads was used for simultaneous processing of up to 48 samples under the same conditions in the development of a simple and rapid procedure to prepare a plant extract for RT reaction. A sandpaper method was also developed by which wood tissue of dormant cuttings could be macerated easily to process with minimal time and effort. It was also demonstrated that the combination use of random primers and oligo dT primer in an RT reaction was efficient for simultaneous cDNA synthesis of viral and viroid RNAs in plant extracts. These template preparation methods were used for the amplification of Grapevine leafroll-associated virus-1,-2, and -3; Grapevine virus A and B; Grapevine rupestris stem pitting-associated virus; Grapevine fleck virus; and Grapevine fanleaf virus. All these viruses tested in this study were reliably detected up to a 10(3)-fold or higher dilution of the original extract. Besides, Hop stunt viroid and Grapevine yellow speckle viroid 1 were well amplified in the same manner as the template preparation and following PCR for virus detection. These methods would con...Continue Reading

References

Oct 1, 1991·The Journal of General Virology·C RitzenthalerL Pinck
Dec 1, 1989·The Journal of General Virology·A M KolunowM A Rezaian
Feb 11, 1988·Nucleic Acids Research·A M Koltunow, M A Rezaian
Jun 1, 1997·Journal of Virological Methods·P La NotteP Saldarelli
Jul 18, 2001·The Journal of General Virology·S SabanadzovicG P Martelli

❮ Previous
Next ❯

Citations

Mar 28, 2008·Archives of Virology·Ryoji Nakaune, Masaaki Nakano
Sep 12, 2008·Archives of Virology·Ryoji NakauneMasaaki Nakano
Jul 6, 2013·Archives of Virology·Takao ItoAkihiko Sato
Nov 17, 2009·Journal of Virological Methods·Esteban A EngelPablo D T Valenzuela
Jan 14, 2009·Plant Physiology and Biochemistry : PPB·Hironori KatohTsutomu Takayanagi
May 25, 2013·The Journal of General Virology·Takao ItoMasaaki Nakano
Mar 21, 2015·Virus Research·Varvara I MaliogkaApostolos D Avgelis
Nov 26, 2011·Journal of Virological Methods·Fatima OsmanAdib Rowhani
Dec 24, 2018·BMC Microbiology·Martin Steen MortensenKaren Angeliki Krogfelt
Jul 7, 2020·EFSA Journal·UNKNOWN EFSA Panel on Plant Health (EFSA PLH Panel)Luisa Rubino

❮ Previous
Next ❯

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.