Development of real-time PCR assay using TaqMan probe for detection and quantification of Rosellinia necatrix in plant and soil

2012 ◽  
Vol 78 (2) ◽  
pp. 115-120 ◽  
Author(s):  
Masahiro Shishido ◽  
Itsuki Kubota ◽  
Hitoshi Nakamura
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Taqman Probe ◽  
Rosellinia Necatrix ◽  
Pcr Assay ◽  
Detection And Quantification

A Diagnostic TaqMan Real-Time PCR Assay for in planta Detection and Quantification of Colletotrichum theobromicola, Causal Agent of Boxwood Dieback

Plant Disease ◽  
2021 ◽  
Author(s):  
Harleen Kaur ◽  
Raghuwinder Singh ◽  
Vinson P. Doyle ◽  
Rodrigo Valverde
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Species Complex ◽  
The United States ◽  
Taqman Probe ◽  
In Planta ◽  
Pcr Assay ◽  
Phytophthora Root Rot ◽  
Specificity And Sensitivity ◽  
Detection And Quantification

Boxwood dieback, caused by Colletotrichum theobromicola, is spreading at an alarming rate in the boxwood industry in the United States. Although C. theobromicola has been accepted as a distinct species within the C. gloeosporioides species complex, it is difficult to distinguish it from other closely related species based on morphology. Moreover, molecular identification of C. theobromicola requires amplification and sequencing of multiple loci, which can be expensive and time consuming. Therefore, a diagnostic TaqMan real-time PCR assay was developed for early and accurate detection and quantification of C. theobromicola in boxwood. The study involved the design of species-specific primers and a TaqMan probe to differentiate C. theobromicola from other closely related Colletotrichum species. The primers and probe discriminate between C. theobromicola and other species in the C. gloeosporioides species complex and can detect C. theobromicola at very low concentrations, illustrating the high specificity and sensitivity of the assay. This TaqMan real-time PCR assay accurately and rapidly distinguishes boxwood dieback from other diseases with similar symptomatology including, Macrophoma blight, Phytophthora root rot, and Volutella blight, as well as some disorders produced by abiotic agents.

A TaqMan real-time PCR assay for detection of Meloidogyne hapla in root galls and in soil

Nematology ◽  
2016 ◽  
Vol 18 (2) ◽  
pp. 147-154 ◽  
Author(s):  
Rumakanta Sapkota ◽  
Andrea M. Skantar ◽  
Mogens Nicolaisen
Keyword(s):  
Disease Management ◽  
Early Detection ◽  
Real Time ◽  
Real Time Pcr ◽  
Taqman Probe ◽  
Meloidogyne Hapla ◽  
Pcr Assay ◽  
Soil Dna ◽  
Dilution Series ◽  
Detection And Quantification

Early detection and quantification of Meloidogyne hapla in soil is essential for effective disease management. The purpose of this study was to develop a real-time PCR assay for detection of M. hapla in soil. Primers and a TaqMan probe were designed for M. hapla detection. The assay detected M. hapla and showed no significant amplification of DNA from non-target nematodes. The assay was able to detect M. hapla in a background of plant and soil DNA. A dilution series of M. hapla eggs in soil showed a high correlation (, ) with Ct values. The assay could predict root-knot development in carrots by testing soils before planting. The assay could be useful for management decisions in carrot cultivation.

scholarly journals Detection of Peanut Allergen Traces with a Real Time PCR Assay - The Challenge to Protect Food-Allergic Consumers

2017 ◽  
Vol 7 (1) ◽  
pp. 32 ◽  
Author(s):  
Dimitra Houhoula ◽  
Stamatios Koussissis ◽  
Vladimiros Lougovois ◽  
John Tsaknis ◽  
Dimitra Kassavita ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Rapid Detection ◽  
Food Products ◽  
Molecular Techniques ◽  
Food Allergens ◽  
Pcr Assay ◽  
Peanut Allergen ◽  
Pcr Method ◽  
Detection And Quantification

The aim of the present study was the implementation of molecular techniques in the detection and quantification of allergic substances of peanut in various kinds of food products, e.g., breakfast cereals, chocolates and biscuits that are frequently related to allergies. In some cases, the presence of peanuts can be due to contamination during production and are not declared on the label. A total of 152 samples were collected from supermarkets and were analysed by a Real Time PCR method. The results indicated that 125 samples (83,3%) were found positive in peanut traces but the most important finding is that from the 84 samples that had no allergen declaration for peanuts, 48 (57,1%) of them were found positive. In conclusion, Real Time PCR can be a very important tool for the rapid detection and quantification of food allergens.

scholarly journals One-step real-time PCR assay for detection and quantification of RNA HCV to monitor patients under treatment in Brazil

2018 ◽  
Vol 22 (5) ◽  
pp. 418-423
Author(s):  
Elisabete Andrade ◽  
Daniele Rocha ◽  
Marcela Fontana-Maurell ◽  
Elaine Costa ◽  
Marisa Ribeiro ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Pcr Assay ◽  
One Step ◽  
Detection And Quantification

scholarly journals Pathogenicity and a TaqMan Real-Time PCR for Specific Detection of Pantoea allii, a Bacterial Pathogen of Onions

Plant Disease ◽  
2019 ◽  
Vol 103 (12) ◽  
pp. 3031-3040 ◽  
Author(s):  
Shabnam Rahimi-Khameneh ◽  
Sanni Hsieh ◽  
Renlin Xu ◽  
Tyler J. Avis ◽  
Sean Li ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Phylogenetic Analyses ◽  
Similarity Index ◽  
Taqman Probe ◽  
Economic Losses ◽  
In Planta ◽  
Pcr Assay ◽  
Blind Test ◽  
Reliable Detection

Bacterial diseases of onion are reported to cause significant economic losses. Pantoea allii Brady, one of the pathogens causing the center rot on onions, has not yet been reported in Canada. We report the pathogenicity of P. allii on commercially available Canadian green onions (scallions). All P. allii-inoculated plants, irrespective of the inoculum concentration, exhibited typical leaf chlorotic discoloration on green onion leaves, which can reduce their marketability. Reisolation of P. allii from infected scallion tissues and reidentification by sequencing and phylogenetic analyses of the leuS gene suggest that the pathogen can survive in infected tissues 21 days after inoculation. This is the first report of P. allii as a potential pathogen of green onions. This study also reports the development and validation of a TaqMan real-time PCR assay targeting the leuS gene for reliable detection of P. allii in pure cultures and in planta. A 642-bp leuS gene fragment was targeted because it showed high nucleotide diversity and positively correlated with genome-based average nucleotide identity with respect to percent similarity index and identity of Pantoea species. The assay specificity was validated using 61 bacterial and fungal strains. Under optimal conditions, the selected primers and FAM-labeled TaqMan probe were specific for the detection of nine reference P. allii strains by real-time PCR. The 52 strains of other Pantoea spp. (n = 25), non-Pantoea spp. (n = 20), and fungi/oomycetes (n = 7) tested negative (no detectable fluorescence). Onion tissues spiked with P. allii, naturally infested onion bulbs, greenhouse infected green onion leaf samples, as well as an interlaboratory blind test were used to validate the assay specificity. The sensitivities of a 1-pg DNA concentration and 30 CFU are comparable to previously reported real-time PCR assays of other bacterial pathogens. The TaqMan real-time PCR assay developed in this study will facilitate reliable detection of P. allii and could be a useful tool for screening onion imports or exports for the presence of this pathogen.

A real time PCR assay for the detection and quantification of orf virus

2006 ◽  
Vol 134 (1-2) ◽  
pp. 140-145 ◽  
Author(s):  
L. Gallina ◽  
F. Dal Pozzo ◽  
C.J. Mc Innes ◽  
G. Cardeti ◽  
A. Guercio ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Orf Virus ◽  
Pcr Assay ◽  
Detection And Quantification
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