Development of the molecular methods for potato virus and viroid detection and prevention

Genome ◽  
1999 ◽  
Vol 42 (4) ◽  
pp. 592-604 ◽  
Author(s):  
Rudra P Singh
Keyword(s):  
Polymerase Chain Reaction ◽  
Developing Countries ◽  
Nucleic Acid ◽  
Potato Virus ◽  
Reverse Transcription ◽  
Virus Detection ◽  
Chain Reaction ◽  
Virus Diseases ◽  
Spot Hybridization ◽  
Polymerase Chain

Potato is the fourth most important food crop in the world and it forms the diet of a billion consumers in developing countries, where potato production is increasing rapidly. However, potato virus diseases in developing countries are one of the major causes of lower yields. Their control requires the development of appropriate virus-detection and seed-production technologies for the region. Recent progress in developing nucleic acid based virus detection methods are reviewed. Refinements of the protocols applicable to the laboratories located in seed producing areas are discussed. Nucleic acid spot hybridization (NASH) and reverse transcription polymerase chain reaction (RT-PCR) methods are described for the detection of viruses and viroids in dormant seed tubers and insect vectors. Although the potato crop is susceptible to over 25 virus and viroid diseases, only universally economically important viruses have been dealt with here. The progress of pathogen-derived resistance for the control of potato virus diseases is elaborated, and the results of field tests indicate their feasibility in virus control.Key words: dot-blot, spot-hybridization, reverse transcription, polymerase chain reaction, transgenic plants.

scholarly journals Specific Detection of Potato Virus A in Dormant Tubers by Reverse-Transcription Polymerase Chain Reaction

Plant Disease ◽  
1998 ◽  
Vol 82 (2) ◽  
pp. 230-234 ◽  
Author(s):  
Rudra P. Singh ◽  
Mathuresh Singh
Keyword(s):  
Polymerase Chain Reaction ◽  
Nucleic Acids ◽  
Potato Virus ◽  
Reverse Transcription ◽  
Rt Pcr ◽  
Specific Detection ◽  
Chain Reaction ◽  
Breeding Lines ◽  
Potato Virus A ◽  
Polymerase Chain

A reverse-transcription polymerase chain reaction (RT-PCR) protocol was developed for the detection of potato virus A (PVA) in dormant tubers. A 255-bp amplified product was produced using a primer pair from the P1 gene of the PVA genome. The 255-bp product was detected in nucleic acids from leaves, tubers, and purified virions and was specific to PVA as determined by Southern blot tests and detection by a PVA-specific probe. When presented with seven potato virus/strain nucleic acids and a viroid, singly and in mixed infections, the primer pair did not amplify any products. Its specificity to PVA was further demonstrated by RT-PCR detection of PVA from the known mixtures of PVA and potato virus Y samples. PVA was detected in foliage nucleic acids at a dilution of 1:1024–1:4096 and tuber nucleic acids at 1:256–1:1024. It was uniformly present in various parts of the potato tuber. PVA was detected in composite tuber samples containing a ratio of infected to healthy sap of 1:29 and was readily detected in tubers of several cultivars or breeding lines, in dormant as well as in sprouting tubers stored at 20–25°C for 4 months.

scholarly journals Reverse transcription-polymerase chain reaction assay for rabies virus detection

2004 ◽  
Vol 56 (3) ◽  
pp. 398-400 ◽  
Author(s):  
J.V. Dantas Junior ◽  
L.M.S. Kimura ◽  
M.S.R. Ferreira ◽  
A.M. Fialho ◽  
M.M.S. Almeida ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Rabies Virus ◽  
Reverse Transcription ◽  
Polymerase Chain Reaction Assay ◽  
Virus Detection ◽  
Chain Reaction ◽  
Polymerase Chain

scholarly journals Nucleic Acid Testing of SARS-CoV-2

2021 ◽  
Vol 22 (11) ◽  
pp. 6150
Author(s):  
Hee-Min Yoo ◽  
Il-Hwan Kim ◽  
Seil Kim
Keyword(s):  
Polymerase Chain Reaction ◽  
Nucleic Acid ◽  
Reverse Transcription ◽  
Diagnostic Methods ◽  
Detection Methods ◽  
Molecular Diagnostic ◽  
Nucleic Acid Testing ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Polymerase Chain

The coronavirus disease 2019 (COVID-19) has caused a large global outbreak. It is accordingly important to develop accurate and rapid diagnostic methods. The polymerase chain reaction (PCR)-based method including reverse transcription-polymerase chain reaction (RT-PCR) is the most widely used assay for the detection of SARS-CoV-2 RNA. Along with the RT-PCR method, digital PCR has emerged as a powerful tool to quantify nucleic acid of the virus with high accuracy and sensitivity. Non-PCR based techniques such as reverse transcription loop-mediated isothermal amplification (RT-LAMP) and reverse transcription recombinase polymerase amplification (RT-RPA) are considered to be rapid and simple nucleic acid detection methods and were reviewed in this paper. Non-conventional molecular diagnostic methods including next-generation sequencing (NGS), CRISPR-based assays and nanotechnology are improving the accuracy and sensitivity of COVID-19 diagnosis. In this review, we also focus on standardization of SARS-CoV-2 nucleic acid testing and the activity of the National Metrology Institutes (NMIs) and highlight resources such as reference materials (RM) that provide the values of specified properties. Finally, we summarize the useful resources for convenient COVID-19 molecular diagnostics.

scholarly journals Ultra-sensitive Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) Antigen Detection for the Diagnosis of Coronavirus Disease 2019 (COVID-19) in Upper Respiratory Samples

2021 ◽  
Author(s):  
Hannah Wang ◽  
Catherine A Hogan ◽  
Michelle Verghese ◽  
Daniel Solis ◽  
Mamdouh Sibai ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Nucleic Acid ◽  
Severe Acute Respiratory Syndrome ◽  
High Throughput ◽  
Reverse Transcription ◽  
Chain Reaction ◽  
Antigen Assay ◽  
Upper Respiratory ◽  
Polymerase Chain ◽  
Respiratory Specimens

Abstract An ultra-sensitive severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) nucleocapsid antigen assay (S-PLEX, MesoScale Diagnostics) was evaluated in 250 retrospective and 200 prospective upper respiratory specimens. In samples with cycle threshold <35, there was 95%–98% positive and 93%–96% negative percent agreement with reverse transcription-polymerase chain reaction. S-PLEX may provide a high-throughput alternative to nucleic acid-based testing for coronavirus disease 2019 (COVID-19) diagnosis.

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